DETAILED ACTION
Final Rejection
Notice of Pre-AIA or AIA Status
1. The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Status of the Claims
2. Claims 1, 3, and 5-20 as amended on 02/18/2026 are pending and are under examination in this office action. Applicant has cancelled claims 2, and 4.
Amended claims 1, 3, and 5-20 are under examination in this office action.
Priority (modified)
3. In view of the applicant’s amendment to the claims and specification filed on 02/18/2026 the instant amended application is denied the earlier priority date of U.S. Provisional Patent Application No. 63/085,004, filed September 29, 2020. The amendment constitutes a new matter and the priority date of introduction of the new matter 02/20/2026 is the new eligible priority date or effective filing date.
There instant application is subjected to a 35 U.S.C. 112(a) new matter rejection as recited further in the office action.
The instant application and the filed published article by Chen et al 2022 as evidence differ by two co-authors Yuxin Pan and Jiawei Zio. Co-inventor of instant application Zhi Zhao and co-authors of the article Yuxin Pan and Jiawei Zio appears to have contributed similarly to the invention “investigation”, however, Yuxin Pan and Jiawei Zio are not co-inventors on the application. Thus, the third party Yuxin Pan and Jiawei Zio are co-authors on the published article.
The published article by one of the co-inventors Chen et al 2022 (Biosensors and Bioelectronics 202 (2022) filed with an Affidavit under 37 C.F.R § 1.132 served as a prior art under 35 U.S.C. 102(a)(1).
Information Disclosure Statement
4. The information disclosure statement (IDS) submitted on 03/03/2022 and 10/30/2024 is in-compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
OBJECTION: Amendments to the Specification
The Applicant amended paragraph [002.1] of the specification to update the sequence listing paragraph in view of the corrected ST.25 compliant sequence listing submitted herewith. Paragraph [002.1] was added to the application in the response to the Non-Final Office Action filed November 21, 2025, and amended in the first supplemental response filed on February 18, 2026. No new matter has been added.
In view of the foregoing amendments and remarks that supplement the response to the Non-Final Office Action filed November 21, 2025, and the first supplemental response filed on February 18, 2026, the Applicant respectfully requests reconsideration and reexamination of this application and the timely allowance of the pending claims. This is believed to be a complete and proper response to the Sequence Listing Report.
In Response: Applicants amendment to the specification incorporating nanobody sGP49 and sGP7 CDR SEQ ID NOs: 1-6 and the research publication by a co-inventor Chen et al 2022 as recited supra has introduced a new matter and thus a new ground of rejection under 35 U.S.C. 112(a), 35 U.S.C. 102(a)(1) and modification of pending 35 U.S.C. 103, and non-statutory double patenting rejection as recited infra. Applicant should consider the New Matter in response to this objection.
Response to Affidavit Declaration Under Rule 37 CFR § 1.132
5. Applicant on 11/21/2025 filed (total pages 68) an affidavit under rule 37 CFR § 1.132 traversing rejections or objections citing the following reasons:
(i) One of the inventors Dr. Chao Wang, Ph.D. who is also a co-author of the published research paper (co-authored by Chen et al 2022 (Biosensors and Bioelectronics, vol 202, 113971) related to the instant invention and signed a declaration under rule 37 CFR § 1.132 that Dr. Chao Wang, and other co-authors were in possession of the amino acid sequences of the CDRs of the sGP49 and sGP7 nanobodies described in the instant claimed application 17/489273.
(ii) The declaration comprises a statement about personal knowledge and contemporaneous laboratory records maintained under Dr. Chao Wang’s supervision.
(iii) Published research paper pdf copy by Chen et al 2022 (Biosensors and Bioelectronics, vol 202, 113971) comprising the amino acid sequences of the CDRs of the sGP49 and sGP7 nanobodies (Fig. S1) and other figures and published supplementary data.
(iv) Dr. Chao Wang claimed or declared that it was an inadvertent omission of the amino acid sequences of the CDRs of the sGP49 and sGP7 nanobodies in the filing of the application specification and sequence listing for the application 17489273 on 09/29/2021 or in the U.S. provisional filing 63/085004 filed on 09/29/2020.
(iv) E-mail from Shoukai Kang printout (dates 08/11/2017) indicating correspondence with collaborators about Nanobody DNA sequences and order for gene synthesis for expression.
(v) E-mail about binding affinity experiment of nanobody (date 04/25/2017).
Applicant’s affidavit-traversing rejections or objections under 37 CFR 1.132 is considered by the examiner but was not found persuasive insofar that it raises “new matter” (see rejection below) and view of at least the following reasons discussed below:
Applicant did not recite in the original specification of the application number 17/489273 (filed on 09/29/2021) or in the U.S. provisional application 63/085004 (filed on 09/29/2020) that applicant has possession of the antibody e.g. there is no evidence (e.g. examples) that the instantly claimed antibodies (camelid/non-camelind) were sequenced or is evidence of possession of the CDR’s of the instantly claimed antibodies e.g. sequencing the CDRs of the claimed sGP7 and sGP49 nanobodies. Additionally, the specification does not indicate applicant possession of a biological deposit of the claimed nanobodies or antibodies at the time of filing.
It is important to note that the amino acid sequences for the CDRs of the sGP7 and sGP49 nanobodies although very important to obtain the patent were placed in the supplementary data Figure S1 (a) and not in the main content of published research paper that could have been obviously visible and accessible to the researchers and inventors in the art. The main part of the article has many figures but not the claimed nanobody CDR sequences.
Additionally, an email or article regarding subject matter, filed prior to the instant filing date, that was “inadvertently” not incorporated into the instant application at the time of filing; does not constitute possession at the time of filing of the instant application of this subject matter.
Therefore, the applicant was clearly not in possession of the claimed sGP7 and sGP49 nanobody CDR sequences at the time of original filing of the application 17/489273 filed on 09/29/2021 or in the U.S. provisional application 63/085004 filed on 09/29/2020.
Claim Rejections - 35 USC § 112, First Para (New)
6. The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
7. The instant claims 1, 3, and 5-20 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
This is a new matter rejection. The instant specification does not have support for the claimed sGP7 nanobody and sGP49 nanobody because there is no recitation of sGP7 and sGP49 nanobody CDRs sequencing and the specification did not encompass the nanobody sGP7 and sGP49 nanobody CDR sequences in the specification and claims filed in the provisional application 63/085,004, filed September 29, 2020, or instant application 17/489,273 on the filing date 09/29/2021. Applicant only describe the names of the nanobodies sGP7 and sGP49 with binding constant kD values. Applicant’s Affidavit under 37 C.F.R § 1.132 comprising the published article Chen et al 2022 (Biosensors and Bioelectronics 202 (2022) and the instant claimed sGP7 and sGP49 nanobody CDR sequences was not a part of original provisional and instant application and specification.
8. The instant claims 1, 3, and 5-20 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claims contain subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
The claims 1, 3, and 5-17 are directed to methods, inter alia, of detecting Ebola virus or Ebola virus glycoproteins or Ebola virus secreted glycoprotein in a sample using gold and or other plasmonic metallic nanoparticle that are conjugated with at least two sets of nanobodies comprising three variable CDRs and nanobody identified as sGP7 (kD 199 nM) binds to second epitope and sGP49 (kD 4.6 nM) binds to first epitope of a secreted glycoprotein (sGP) of Ebola virus. The claims 1, 3, and 5-20 of the invention require use of gold/plasmonic metallic conjugated with sGP7 and sGP49 nanobody that bind to Ebola virus secreted glycoproteins in samples or standard purified Ebola virus secreted glycoproteins.
The claims 1, 3, and 5-20 and the entire specification and figure do not provide the sequences of the three CDRs of the sGP7 and sGP49 nanobody at the effective filing date of the claimed invention. Additionally, the scope of the instant claims (particularly claim 1) are “broad” in scope encompassing camelid and non-camelid antibodies that contain the CDR’s but are not limited in the protein structure of the remaining antibody portion which may influence the functional integrity of the antibody e.g. structure effects function.
Additionally, the claims and specification do not provide the sequence of the first and second epitope of Ebola virus secreted glycoproteins to which the nanobody sGP7 and sGP49 are claimed to bind. One of the ordinary skills in the art is unable to envision the location, the sequence or a structure of first and second epitope of Ebola virus secreted glycoproteins.
Further, claims 18-20 are directed to a device and a system, without recitation of structure, but merely contain the assay components of claim 1. In this regard, consistent with the specification disclosure, conventional apparatuses comprising “reaction chambers” and systems further containing means of detecting antibody (nanobody) antigen binding were known in the prior art. See e.g. Ye et al. March 2020: 10(10);4359-4373 and prior art cited therein.
The claims 1, 3, and 5-20 and the entire specification and figure do not provide the sequences of the three CDRs of the sGP7 and sGP49 nanobody at the effective filing date of the claimed invention. The claims and specification do not provide the sequence of the first and second epitope of Ebola virus secreted glycoproteins to which the nanobody sGP7 and sGP49 are claimed to bind.
One of the ordinary skills in the art is unable to envision the location, the sequence or a structure of first and second epitope of Ebola virus secreted glycoproteins.
The para [011] of specification describes, in some embodiments, a given antibody, or antigen binding portions thereof, comprises a mass of between about 10 kDa and about 20 kDa (e.g., about 11 kDa, about 12 kDa, about 13 kDa, about 14 kDa, about 15 kDa, about 16 kDa, about 17 kDa, about 18 kDa, or about 19 kDa). The recited molecular weights are generic for antibodies or antigen binding portions of antibodies and does not determine specificity of antigenic epitope and antibody binding. The recited molecular weights do not provide description on structure of antibodies that confer specificity to the claimed antibodies and hence does not fulfill the requirement of written description of the antibodies claimed in invention.
In the instant application, the applicant is claiming a method that uses sGP7 and sGP49 nanobody for immunologically detecting an Ebola virus secreted glycoprotein, the method comprising detecting an Ebola virus secreted glycoprotein in a specimen, from a subject, that involves interaction and formation of the antigen or epitope and nanobody complex. There is no specific description of structure of the antibodies as required by Amgen (When an antibody is claimed, 35 U.S.C. § 112(a) requires adequate written description of the antibody itself). The description of structure of the claimed antibodies/monoclonal antibodies/scFV, antigen binding portions of antibodies/nanobodies that is required and is not found in the specification is as below:
(a). The nucleotide or amino acid sequence of variable region of heavy and variable region of light chain that confers the specificity to the antibodies/monoclonal antibodies or alternative forms of antibodies to bind to a specific amino acid region epitope of an Ebola virus secreted glycoprotein.
(b). The nucleotide or amino acid sequence of hypervariable domains of heavy (H1-H3) and variable light chain (L1-L3) called Complementarity-Determining Regions (CDRs) that confers the specificity to the antibodies/monoclonal antibodies or alternative forms of antibodies to bind to a specific amino acid region epitope of an Ebola virus secreted glycoprotein.
The recited claims or specification, drawings and figures does not provide a description of the specific nucleotide or amino acid structure of the claimed antibodies or alternative forms of antibodies that binds to the claimed Ebola virus glycoprotein, and thus raises doubt as to possession of the claimed invention at the time of effective filing date of the invention.
In the context of claimed invention involving nanobody sGP7 and sGP49 that bind an Ebola virus secreted glycoprotein, the U.S. Court of Appeals for the Federal Circuit (Federal Circuit) recently decided Amgen v. Sanofi, 872 F.3d 1367 (Fed. Cir. 2017), which concerned adequate written description for claims drawn to antibodies.
The Federal Circuit explained in Amgen that when an antibody is claimed, 35 U.S.C. § 112(a) requires adequate written description of the antibody itself. Amgen, 872 F.3d at 1378-79. The Amgen court expressly stated that the so-called "newly characterized antigen" test should not be used in determining whether there is adequate written description under 35 U.S.C. § 112(a) for a claim drawn to an antibody. Citing its decision in Ariad Pharmaceuticals, Inc. v. Eli Lilly & Co., the court also stressed that the "newly characterized antigen" test could not stand because it contradicted the quid pro quo of the patent system whereby one must describe an invention in order to obtain a patent. Amgen, 872 F.3d at 1378-79, quoting Ariad Pharmaceuticals, Inc. v. Eli Lilly & Co., 598 F.3d 1336, 1345 (Fed. Cir. 2010). In view of the Amgen decision, adequate written description of a newly characterized antigen alone is not considered adequate written description of a claimed antibody to that newly characterized antigen, even when preparation of such an antibody is routine and conventional. Id. See Amgen Inc. v. Sanofi, 872 F.3d 1367, 1378, 124 USPQ2d 1354, 1361 (Fed. Cir. 2017) (“knowledge of the chemical structure of an antigen [does not give] the required kind of structure-identifying information about the corresponding antibodies”); see also Centocor Ortho Biotech, Inc. v. Abbott Labs., 636 F.3d 1341, 1351-52, 97 USPQ2d 1870, 1877 (Fed. Cir. 2011)(patent disclosed the antigen the claimed antibody was supposed to bind, but did not disclose any antibodies with the specific claimed properties). In the instant claims and specification, the sequence of the epitopes or antigenic determinants of the secreted glycoprotein of an Ebola virus to which the claimed sGP7 and sGP49 nanobody binds are not disclosed at the effective filing date of the claimed invention. Therefore, one of the ordinary skills in the art raises a doubt that whether the applicant had possession of the claimed sGP7 and sGP49 nanobody at the effective filing date of the claimed invention.
9. Claims 1, 3, and 5-20 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the enablement requirement (deposit rejection). Claims 1, 3, and 5-20 contain subject matter which was not described in the specification in such a way as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention.
At the original filing date of the application 17/489273 (09/29/2021) or the US provisional application 63/085004 (09/29/2020) the claims and specification lacked the sequences for CDRs of the nanobody sGP7 and nanobody sGP49 that was a required element of the claimed invention. As a required elements it must be known and readily available to the public or obtainable by a repeatable method set forth in the specification, or otherwise readily available to the public. If it is not so obtainable or available, the enablement requirements of 35 U.S.C. § 112, first paragraph, may be satisfied by a deposit of the claimed nanobody sGP7 and nanobody sGP49 or more specifically the recombinant DNA plasmid expression clones for producing the claimed nanobody sGP7 and nanobody sGP49. See 37 CFR 1.802. One cannot practice the claimed invention without the claimed nanobody sGP7 and nanobody sGP49. Therefore, access to the claimed nanobody sGP7 and nanobody sGP49 is required to practice the invention. The specification does not provide a repeatable method for readily obtaining or producing the claimed nanobody sGP7 and nanobody sGP49 without access to the claimed recombinant DNA plasmid expression clones and it does not appear to be a readily available material. The production of nanobodies by phage display as recited in the specification, although is known in the art, the probability of obtaining the claimed nanobody without reciting the sequence is uncertain and selection of recombinant clones producing nanobody binding to specific epitopes of sGP of Ebola virus is a laborious process.
Deposit of the claimed recombinant DNA plasmid expression clones for producing the claimed nanobody sGP7 and nanobody sGP49 in a recognized deposit facility would satisfy the enablement requirements of 35 U.S.C. 112, because the claimed first antibody would be readily available to the public to practice the invention claimed, see 37 CFR 1.801- 37 CFR 1.809.
If the deposit was made under the terms of the Budapest Treaty, then a statement, affidavit or declaration by applicants, or a statement by an attorney of record over his or her signature and registration number, or someone empowered to make such a statement, stating that all restrictions imposed by the depositor on the availability to the public of the deposited material will be irrevocably removed upon the granting of the patent, would satisfy the deposit requirement.
If the deposit was not made under the Budapest Treaty, then in order to certify that the deposit meets the criteria set forth in 37 CRF 1.801-1.809 and MPEP 2402-2411.05, applicants may provide assurance of compliance by statement, affidavit or declaration or by someone empowered to make the same or by a statement by an attorney of record over his or her signature and registration number showing that:
(a) during the pendency of the application, access to the invention will be afforded to the Commissioner upon request;
(b) all restrictions upon availability to the public will be irrevocably removed upon granting of the patent;
(c) the deposit will be maintained in a public depository for a period of 30 years or 5 years after the last request or for the enforceable life of the patent, whichever is longer;
(d) a test of the viability of the biological material at the time of deposit (see 37 CFR 1.807); and
(e) the deposit will be replaced if it should ever become inviable.
In addition, the identifying information set forth in 37 CFR 1.809(d) should be added to the specification. See 37 CFR 1.803 - 37 CFR 1.809 for additional explanation of these requirements.
Based on the reasons recited supra, one of the ordinary skilled in the art could not make or use the invention defined in and commensurate with the claims without access to the specific biological material. See MPEP § 2164.06(a).
To satisfy the enablement requirement a deposit must be made “prior to issue” but need not be made prior to filing the application. In reLundak, 773 F.2d 1216, 1223, 227 USPQ 90, 95 (Fed. Cir. 1985).
Thus, the pending claims 1, 3, and 5-20 are not enabled for the full scope of the use of the claimed invention due to non-compliance with deposit requirement as recited supra.
Response to Applicant’s Arguments
Applicant’s arguments filed on 11/21/2025 with respect to claims 1, 3, and 5-20 have been considered but are moot because the new matter and new ground of rejection does not rely on only prior applied reference but also include new references as prior arts of record for any teaching or matter specifically challenged in the argument.
Applicant’s argument:
Rejections Under 35 U.S.C. § 112 Claims 1, 3 and 5-20 are rejected under 35 U.S.C. § 112(a) or 35 U.S.C. § 112 (pre-AIA ), first paragraph, as allegedly failing to comply with the enablement requirement. While the Applicant respectfully disagrees with the Examiner's assertion that the claims as written lack enablement, the Applicant herewith discloses the sequences of the CDRs of the 7 sequences is shown in Figure S1(a) of the Supplementary Information for Chen et al., "Synthetic nanobody- functionalized nanoparticles for accelerated development of rapid, accessible detection of viral antigens," Biosensors and Bioelectronics 202 (2022) 113971 of which each of the inventors is listed as an author and which publication is attached hereto as Exhibit 1. Thus, as noted above, the Applicant has further fulfilled the Enablement Requirement by herewith disclosing the sequences of each of the CDRs of the nanobodies recited in the claims. Accordingly, the Applicant respectfully requests that this rejection be withdrawn. Conclusion In view of the foregoing amendments and remarks, Applicant respectfully requests reconsideration and reexamination of this application and the timely allowance of the pending claims.
In Response: A research paper co-authored by Chen et al (2022) filed on 06/18/2025 disclosing sGP and sGP47 nanobody CDR1-3 sequences in Fig S1 (a) of the supplementary page 6 are published after the effective filing date of the claimed inventions. The sequences are not in computer readable format files and are without SEQ ID NO and additionally SEQ ID NO are not recited in the claims. See, For Sequence Listing Format MPEP 2400 et seq.
Applicant’s arguments filed on 06/18/2025 with respect to claims 1, 3, and 5-20 have been considered but are not persuasive and the rejections are maintained.
Claim Rejections - 35 USC § 112(b) (New)
10. The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 1, 3 and 5-20 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
The instant claims 1, 3, 18 and 19 contains the trademark/trade name “nanobody” (nanobody also reads on plural nanobodies). "NANOBODY" is a registered trademark of ABLYNX N.V. It was filed with the USPTO on March 19, 2012. Where a trademark or trade name is used in a claim as a limitation to identify or describe a particular material or product, the claim does not comply with the requirements of 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph. See Ex parte Simpson, 218 USPQ 1020 (Bd. App. 1982). The claim scope is uncertain since the trademark or trade name cannot be used properly to identify any particular material or product. A trademark or trade name is used to identify a source of goods, and not the goods themselves. Thus, a trademark or trade name does not identify or describe the goods associated with the trademark or trade name. In the present case, the trademark/trade name is used to identify/describe “single domain-antibodies (sdAbs)” or “VHH antibodies”, and “these are antibody fragments derived from heavy-chain only IgG antibodies found in the Camelidae family”, and, accordingly, the identification/description is indefinite. (See, evidence by Jin et al 2023 (Int J Mol Sci. 2023 Mar 22;24(6):5994), abstract).
Claim Rejections - 35 USC § 102 (New)
11. In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
Claims 1, 5-7, 9-10, 12 and 14-20: The instant claims 1, 5-7, 9-10, 12 and 14-20 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971, Combined PDF printout of entire article page 1-10 and associated supplementary data on figures, tables and methods page 1-40).
Claim 1: Chen et al 2022 anticipated instant claim 1 by disclosing the instant claimed method of detecting Ebola virus in a sample (See, abstract, page 2, materials and methods) the method comprising: contacting the sample with a plurality of gold nanoparticles (AuNPs) (See, page 3 col 1 section 2.3) and/or other plasmonic metal nanoparticles (MNPs) that are conjugated with at least two sets of nanobodies that each comprise a universal scaffold (See, page 3 col 2 para 1 lines 11-16) and three variable complementarity-determining regions, wherein at least a first set of nanobodies each comprise a mono-binder sGP49 nanobody having a kD of about 4.6 nM that binds to a first epitope of a secreted glycoprotein (sGP) from the Ebola virus, wherein complementarity-determining regions (CDRs) of the sGP49 nanobody comprise the amino acid sequences of SEQ ID NOs:4-6, and wherein at least a second set of nanobodies each comprise a mono-binder sGP7 nanobody having a kD of about 199 nM that binds to a second epitope of the sGP from the Ebola virus, wherein CDRs of the sGP7 nanobody comprise the amino acid sequences of SEQ ID NOs:1-3, under conditions sufficient for the first and second sets of nanobodies to bind to the first or second epitopes of the sGP from the Ebola virus in the sample to produce bound sGPs; or contacting the sample with a plurality of gold nanoparticles (AuNPs) and/or other plasmonic metal nanoparticles (MNPs) that are conjugated with a single set of nanobodies that each comprise a universal scaffold and three variable complementarity-determining regions that bind to an identical epitope from different monomers of a dimerized secreted glycoprotein (sGP) from the Ebola virus under conditions sufficient for the nanobodies to bind to the identical epitope from the different monomers of the dimerized sGP from the Ebola virus in the sample to produce bound sGPs, wherein the single set of nanobodies each comprise a mono-binder sGP49 nanobody having a kD of about 4.6 nM (See, page 8, para 1, and Fig 2), wherein complementarity-determining regions (CDRs) of the sGP49 nanobody comprise the amino acid sequences of SEQ ID NOs:4-6 (See, supplementary Fig S1 Fig S1 on page supplementary 6), or a mono-binder sGP7 nanobody having a kD of about 199 nM (See, page 8, para 1, and Fig 2), wherein CDRs of the sGP7 nanobody comprise the amino acid sequences of SEQ ID NOs:1-3 (See, supplementary Fig S1 on page supplementary 6); and, detecting the sGPs from the Ebola virus when one or more aggregations of the bound sGPs form with one another, thereby detecting the Ebola virus in the sample (See, abstract, methods, page 3, col 1 , 2.3 Antigen detection; Fig 1-4, figures, tables, results, entire article including supplementary data and results and all supplementary figures, See, Workflow Fig S15 on suppl page 26, Fig S21 suppl page 30).
Chen et al 2022 anticipated sGP7 nanobody CDR1, CDR2 and CDR3 amino acid sequences as shown below:
Qy: Instant SEQ ID NO: 1, Db: sGP7 CDR1 by Chen et al 2022, supplementary page 6 Fig. S1 (a).
Query Match 100.0%; Best Local Similarity 100.0%;
Qy 1 FSSSNYA 7
|||||||
Db 1 FSSSNYA 7
Qy: Instant SEQ ID NO: 2, Db: sGP7 CDR2 by Chen et al 2022, supplementary page 6 Fig. S1 (a).
Query Match 100.0%; Best Local Similarity 100.0%;
Qy 1 RWDNVKA 7
|||||||
Db 1 RWDNVKA 7
Qy: Instant SEQ ID NO: 3, Db: sGP7 CDR3 by Chen et al 2022, supplementary page 6 Fig. S1 (a).
Query Match 100.0%; Best Local Similarity 100.0%;
Qy 1 MVDDYWDPG 9
|||||||||
Db 1 MVDDYWDPG 9
Chen et al 2022 anticipated sGP49 nanobody CDR1, CDR2, and CDR3 amino acid sequences as shown below: Qy: Instant SEQ ID NO: 4, Db: sGP49 CDR1 by Chen et al 2022, supplementary page 6 Fig. S1 (a). Query Match 100.0%; Best Local Similarity 100.0%;
Qy 1 DTYYIYA 7
|||||||
Db 1 DTYYIYA 7
Qy: Instant SEQ ID NO: 5, Db: sGP49 CDR2 by Chen et al 2022, supplementary page 6 Fig. S1 (a).
Query Match 100.0%; Best Local Similarity 100.0%;
Qy 1 RSSSAAD 7
|||||||
Db 1 RSSSAAD 7
Qy: Instant SEQ ID NO: 6, Db: sGP49 CDR3 by Chen et al 2022, supplementary page 6 Fig. S1 (a).
Query Match 100.0%; Best Local Similarity 100.0%;
Qy 1 SWHVRFDDYEKVISMQEV 18
||||||||||||||||||
Db 1 SWHVRFDDYEKVISMQEV 18
Chen et al 2022 anticipated instant claim 1 as recited supra.
Claim 5. Chen et al 2022 anticipated the method of claim 1, wherein the detection step comprises determining a change in absorbance at a resonance wavelength of the AuNPs and/or the other MNPs (See, Chen et al 2022 page 5, section 3.3, the combined PDF article Supplementary data page 14, Fig S7 and legends).
Claim 6. Chen et al 2022 anticipated the method of claim 1, comprising quantifying an amount of the sGPs and/or the Ebola virus in the sample (See, Chen et al 2022, page 2 Fig 1 and legends, page 9, col 1 section 4. conclusion, the combined PDF article Supplementary data page 13, last 2 para).
Claim 7. Chen et al 2022 anticipated the method of claim 1, further comprising centrifuging the aggregations of the bound sGPs prior to and/or during the detecting step (See, Fig S11 and legends, page 6 table 1 col 1).
Claim 9. Chen et al 2022 anticipated the method of claim 1, comprising drop casting the aggregations of the bound sGPs prior to the detecting step (See, Figure S6-S8, S10 and associated legends, section 4.5 para 1, supplementary page 16).
Claim 10. Chen et al 2022 anticipated the method of claim 1, comprising obtaining the sample from a subject (See, abstract, human serum, page 7 col 1 section 3.6 whole blood (is obtained from a subject)).
Claim 12. Chen et al 2022 anticipated the method of claim 10, comprising detecting the Ebola virus within about 20 minutes or less of obtaining the sample from the subject (See, abstract, page 3 section 3.2, Figure S11 legends, suppl page 23 section 7.2 for 20 minutes).
Claim 14. Chen et al 2022 anticipated the method of claim 10, wherein the sample comprises blood, plasma, serum, saliva, sputum, or urine (See, human plasma, Table S4, column Diagnostic Sensitivity, row 1, page 7 col 1 section 3.6)
Claim 15. Chen et al 2022 anticipated the method of claim 1, wherein the detecting step comprises measuring a colorimetric change when the one or more aggregations of the bound sGPs form with one another (See, page 1 Fig 1 and legend, col 2 para 1, page 4 col 1 section 3.3 colorimetric sensing of sGP).
Claim 16. Chen et al 2022 anticipated the method of claim 15, comprising visually detecting the colorimetric change when the one or more aggregations of the bound sGPs form with one another (See, Page 1, Fig 1 and legends, page 4, col 1 section 3.3, page 6 Table 1).
Claim 17. The method of claim 15, comprising detecting the colorimetric change when the one or more aggregations of the bound sGPs form with one another using a spectrometer (See, Page 4, col 1-2 section 3.3, page 6 Table 1, Suppl page 23 section 7.2).
Claim 18. Chen et al 2022 anticipated the instant claim 18, a device, comprising at least one reaction chamber (See, page 2, Fig 1 (d) and legends, page 4 col 1 para 1 device, page 6 col 2 section 3.5 device (Nano2RED), Suppl page 23 section 7.2 portable spectrophotometer as device, Figure S3) or substrate comprising a plurality of gold nanoparticles (AuNPs) and/or other plasmonic metal nanoparticles (MNPs) that are (i) conjugated with at least two sets of nanobodies that each comprise a universal scaffold and three variable complementarity-determining regions, wherein at least a first set of nanobodies each comprise a mono-binder sGP49 nanobody having a kD of about 4.6 nM that binds to a first epitope of a secreted glycoprotein (sGP) from an Ebola virus, wherein complementarity-determining regions (CDRs) of the sGP49 nanobody comprise the amino acid sequences of SEQ ID NOs:4-6, and wherein at least a second set of nanobodies each comprise a mono-binder sGP7 nanobody having a kD of about 199 nM that binds to a second epitope of the sGP from the Ebola virus, wherein CDRs of the sGP7 nanobody comprise the amino acid sequences of SEQ ID NOs:1-3, when the reaction chamber or substrate receives a sample that comprises the Ebola virus under conditions sufficient for the first and second sets of nanobodies to bind to the first or second epitopes of the sGP from the Ebola virus in the sample to produce bound sGPs and one or more aggregations of the bound sGPs to produce a colorimetric change in the reaction chamber or on the substrate; or (ii) conjugated with a single set of nanobodies that each comprise a universal scaffold and three variable complementarity-determining regions that bind to an identical epitope from different monomers of a dimerized secreted glycoprotein (sGP) from the Ebola virus under conditions sufficient for the nanobodies to bind to the identical epitope from the different monomers of the dimerized sGP from the Ebola virus in the sample to produce bound sGPs and one or more aggregations of the bound sGPs to produce a colorimetric change in the reaction chamber or on the substrate, wherein the single set of nanobodies each comprise a mono-binder sGP49 nanobody having a kD of about 4.6 nM, wherein complementarity- determining regions (CDRs) of the sGP49 nanobody comprise the amino acid sequences of SEQ ID NOs:4-6, or a mono-binder sGP7 nanobody having a kDof about 199 nM, wherein CDRs of the sGP7 nanobody comprise the amino acid sequences of SEQ ID NOs:1-3 (See, entire article including supplementary data by Chen et al 2022 and rejection recited for claim 1 for SEQ ID NO: 1-6, See, Workflow Fig S15 on suppl page 26, Fig S21 suppl page 30).
Claim 19. Chen et al 2022 anticipated instant claim 19 a system (See, abstract, page 3 col 1 section 2.5, Fig 4 i-k and legends, Fig S13, Suppl page 8 section 3.1, Fig S3), comprising: a device, comprising at least one reaction chamber or substrate comprising a plurality of gold nanoparticles (AuNPs) and/or other plasmonic metal nanoparticles (MNPs) that are (i) conjugated with at least two sets of nanobodies that each comprise a universal scaffold and three variable complementarity-determining regions, wherein at least a first set of nanobodies each comprise a mono-binder sGP49 nanobody having a kD of about 4.6 nM that binds to a first epitope of a secreted glycoprotein (sGP) from an Ebola virus, wherein complementarity-determining regions (CDRs) of the sGP49 nanobody comprise the amino acid sequences of SEQ ID NOs:4-6, and wherein at least a second set of nanobodies each comprise a mono-binder sGP7 nanobody having a kD of about 199 nM that binds to a second epitope of the sGP from the Ebola virus, wherein CDRs of the sGP7 nanobody comprise the amino acid sequences of SEQ ID NOs:1-3, when the reaction chamber or substrate receives a sample that comprises the Ebola virus under conditions sufficient for the first and second sets of nanobodies to bind to the first or second epitopes of the sGP from the Ebola virus in the sample to produce bound sGPs and one or more aggregations of the bound sGPs to produce a colorimetric change in the reaction chamber or on the substrate; or (ii) conjugated with a single set of nanobodies that each comprise a universal scaffold and three variable complementarity-determining regions that bind to an identical epitope from different monomers of a dimerized secreted glycoprotein (sGP) from the Ebola virus under conditions sufficient for the nanobodies to bind to the identical epitope from the different monomers of the dimerized sGP from the Ebola virus in the sample to produce bound sGPs and one or more aggregations of the bound sGPs to produce a colorimetric change in the reaction chamber or on the substrate, wherein the single set of nanobodies each comprise a mono-binder sGP49 nanobody having a kDof about 4.6 nM, wherein complementarity-determining regions (CDRs) of the sGP49 nanobody comprise the amino acid sequences of SEQ ID NOs:4-6, or a mono-binder sGP7 nanobody having a kDof about 199 nM, wherein CDRs of the sGP7 nanobody comprise the amino acid sequences of SEQ ID NOs:1-3; and, an electromagnetic radiation detection apparatus positioned, or positionable, within sufficient proximity to the device to detect one or more colorimetric changes produced in or on the reaction chamber or substrate when one or more aggregations of the bound sGPs form with one another in or on the reaction chamber or substrate (See, entire article including supplementary data by Chen et al 2022 and rejection recited for claim 1 for SEQ ID NO: 1-6, See, Workflow Fig S15 on suppl page 26, Fig S21 suppl page 30).
Claim 20. Chen et al 2022 anticipated instant claim 20 the system of claim 19, wherein the electromagnetic radiation detection apparatus comprises a spectrometer; wherein the electromagnetic radiation detection apparatus comprises a microscope; and/or wherein the electromagnetic radiation detection apparatus comprises a light-emitting diode (LED) that transmits light into and/or through the reaction chamber or substrate and a photodetector that detects light from the reaction chamber or substrate (See, entire article including supplementary data by Chen et al 2022 and page 4 col 1-2 section 3.3, page 7 Fig 4, suppl page 23 section 7.2, Suppl page 8 Figure S3 and section 3.1, Figure S13, See, Workflow Fig S15 on suppl page 26, Fig S21 suppl page 30).
Claim Rejections - 35 USC § 103 (Modified)
12. In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or non-obviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
13: Claims 1, 5-7, 9-10, 12 and 14-20: The instant claims 1, 5-7, 9-10, 12 and 14-20 are rejected under 35 U.S.C. 103 as being unpatentable over Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971, Combined PDF printout of entire article page 1-10 and associated supplementary data on figures, tables and methods page 1-40).
Claims 1, 5-7, 9-10, 12 and 14-20: The disclosures of Chen et al 2022 as recited supra are incorporated here in entirety to render obvious the instant claims 1, 5-7, 9-10, 12 and 14-20 under 35 U.S.C. 103.
14. Claims 1, 3, 5-6, 10, 14 are rejected under 35 U.S.C. 103 as being unpatentable over Gehrke et al 2018 (US20180372755A1 published 27 December 2018), and further in view of Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971, Combined PDF printout of entire article page 1-10 and associated supplementary data on figures, tables and methods page 1-40), Liu et al 2017 (published in Microb Cell Fact (2017) 16:223), Ning et al 2017 (Virologica Sinica, 2017, 32 (1), p. 3–15), Zhang et al 2016 (Sci Rep 6, 25856, p. 1-15), Bornholdt et al 2016 (Science. 2016 Mar 4;351(6277):1078-83), Szunerits et al 2020 (Nanomaterials (Basel). 2020 Jun 29;10(7):1271), Wu et al 2020 (ACS Applied Nano Materials, 3(10), 9560-9580), Hung et al 2014 (Nanomedicine. 2014 May;10(4):819-29), Alejo-CanchoIet al 2020 (PLoS Negl Trop Dis. 2020 Feb 18;14(2)), Koh et al 2020 (Sensors 2009, 9, 8130-8145).
Claims 1, 3, 10, 14: Gehrke et al 2018 is in the art and is directed to a method of detecting Ebola virus comprising contacting the biological sample with one or more antibodies directed to epitope on Ebola virus glycoprotein or secreted glycoprotein that is detected by antibody binding to an antigenic site (epitope) wherein one or more detection antibodies are labelled with a gold/silver or metallic nanoparticles and the bound glycoprotein antigen-gold nanoparticle conjugated antibody immunocomplexes are detected. Gehrke et al 2018 teaches additionally, we demonstrated that it could be applied to detect Ebola virus biomarker glycoprotein 1 (GP1) using Marburg anti-GP antibodies (para [0119]). (See, claim 1, claim 3, claim 7, claim 9, claim 11; paragraphs [0008], [0009], [0010], [93-95], [99, 101, 107, 108] [0119], [0120], [0162]. Gehrke et al discloses obtaining a sample of biological origin, or a sample derived from the sample of biological origin, preferably from human patient. The biological samples include, but are not limited to, blood, plasma, serum, saliva, cerebral spinal fluid, ……, sputum, urine, stool, tear, ….., respiratory, intestinal, or genitourinary tract, organ, cell culture, cell culture constituent, tissue sample, tissue section, whole cell, cell constituent, or cell smear (See, paragraph [0110] and claims 1, 7-9).
Gehrke et al 2018 does not teach nanobody that bind to an Ebola virus secreted glycoprotein (sGP).
Chen et al 2022 as recited supra under 35 U.S.C. 102(a)(1) rejection teaches sGP7 and sGP49 nanobody CDR1, CDR2 and CDR3 amino acid sequences SEQ ID NO: 1-3 (sGP7 binding nanobody), and SEQ ID NO: 4-6 (sGP49 binding nanobody) as recited supra with 100% amino acid identity and are incorporated here in entirety.
Liu et al 2017 teaches camelid (llama) nanobodies that specifically bind to an Ebola virus glycoprotein, disclosing characterization of initially selected 10 sdAbs 10 (See, page 3, column 1 for section on characterization of initially selected sdAbs) table 1 listing binding affinity of sdAs nanobodies EBOV-GP-G6, EBOV-GP-H7, EBOV-GP-A8, EBOV-GP-B11, EBOV-GP-G3, EBOV-GP-C11, EBOV-GP-G3, EBOV-GP-B5, and EBOV-GP-D1 bind to epitopes on Ebola virus GP (Table 1, Table 3); and detection of Ebolavirus glycoprotein using the nanobodies. Our results suggest that the selected GP binders also bind the heavily glycosylated GPs with the size more than 100 kDa in purified recombinant GP. (See, Liu et al 2017, additional file 1: Figure S6; page 7; also see title and abstract, entire article, page 8, col 1 para 1, Fig 4-6, entire article).
The claimed nanobody sGP7 binding to the first epitope of sGP has kD 199 nM and sGP49 binding to the second epitope of sGP has kD 4.6. Liu et al teaches thermal stabilization optimization of kD for camelid (llama) sdAb by disclosing three of the GP binding sdAbs showing the highest melting temperatures and/ or best affinities were subject to mutagenesis to improve their stability. Melting temperature and refolding ability were measured by CD (See, Table 3, entire article). Optimizing sdAb kD involves combining affinity maturation techniques, targeted engineering of CDRs and FRs, and comprehensive characterization depending on the specific target and application. As a practical matter, the Patent Office is not equipped to manufacture products by the myriad of processes put before it and then obtain prior art products and make physical comparisons therewith.” In re Brown, 459 F.2d 531, 535, 173 USPQ 685, 688 (CCPA 1972). See e.g. MPEP 2113.
Ning et al 2017 teaches processing of Ebolavirus GP in infected cell (Fig 1) as shown below.
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Ning et al 2017 disclosed above recited Figure 1, See legends on page 5 of the research paper.
The sGP homodimer is 110 kD (page 8, col 1, para 1). GP1, shed GP and ssGP comprise the glycosylated portions of sGP as seen in figure. As recited supra, Liu et al 2017 teaches that selected nanobodies, the GP binders also bind the heavily glycosylated GPs with the size more than 100 kDa in purified recombinant GP. Thus, Liu et al 2017 teaches the nanobodies that bind to sGP of Ebola virus.
Zhang et al 2016 and Bornholdt et al 2016 as recited below teaches monoclonal antibodies that strongly binds to sGP of Ebola virus and is a functional equivalent alternative for the claimed nanobody sGP7 and sGP49 to develop the claimed methods and the inventions.
Zhang et al 2016 teaches Ebola virus monoclonal antibodies that strongly binds to c13C6, Q206, Q314 and Q411 to sGP indicating their epitopes are located in the N-terminus of GP1 subunit (See, page 2 section on Binding activity against pseudo and live Ebola virus infection; and page 3 Fig 1 a - binding).
Bornholdt et al 2016 discloses isolation and characterization of 349 GP-specific monoclonal antibodies (mAbs) from the peripheral B cells of a convalescent donor who survived the 2014 EBOV Zaire outbreak (See, abstract) and 59% of the mAbs reacted to sGP (See, page 1079-Fig 1 C, page 1080-Fig 2, epitope mapping to sGP, ELISA binding).
Szunerits et al 2020 is in the virology and viral diagnostic art and reviews applications of superparamagnetic nanoparticles (MPs) for virus or viral antigen (page 4, Fig 1, e.g. HSV-1 Magnetic particle aggregation assay) detection exploiting magnetic bead-based viral aggregation assays and their integration into different biosensing strategies as they can be easily separated from a complex matrix containing the virus through the application of an external magnetic field (See, abstract, Figure 3, a, b).
Wu et al 2020 is in the viral diagnostic art and reviews teaches magnetic-nanosensor-based virus and pathogen detection strategies before and during COVID-19 using magnetic nanosensing techniques including magnetoresistance, magnetic particle spectroscopy, and nuclear magnetic resonance and magnetic point-of-care diagnostic kits (See, Wu et al 2020, entire article).
Hung et al 2014 is in the virology diagnostic art and teaches magnetic nanoparticle-based immunoassay for rapid detection of influenza infections by using an integrated microfluidic system (See, abstract, Fig 1-6, entire article).
Alejo-Cancho et al 2020 is in the virology diagnostic art and teaches evaluation of a novel microfluidic immunomagnetic agglutination assay method for detection of dengue virus NS1 antigen (See, entire article, Fig 2).
Koh et al 2020 is in the viral diagnostic art and reviews magnetic nanoparticle sensors (See, abstract and entire review).
It would have been obvious to one of the ordinary skill in the art before the effective filing date of the claimed invention to modify the prior art teachings on the method of detection of Ebola virus GP1 glycoprotein taught by Gehrke et al and incorporate the teachings of Chen et al 2022, Liu et al on nanobody binding to heavily glycosylated Ebola virus glycoprotein and additional prior arts teachings including Ning et al to arrive at the invention of claim 1. Ning et al, as recited supra, teaches that the sGP homodimer is 110 kD GP1, shed GP and ssGP comprise the glycosylated portions of sGP and thus the nanobodies disclosed by Liu et al binds to sGP of Ebola virus. The motivation would have been to exploit the properties of nanobodies (SdAbs) such as the smallest naturally occurring antigen binding domains, superior thermal stability, ability to recognize hidden epitopes combined with specificity and binding affinity comparable to conventional antibodies (Abs) and amenable to protein engineering to improve their properties (See, Liu et al, page 1, introduction, para 1). As Liu et al (See, page 3, table 1) has demonstrated specific binding of 10 different nanobodies to an Ebola virus glycoprotein, using the nanobodies. There would have been a reasonable expectation of success to arrive at the invention of instant claim 1 based on the applied prior art teachings.
Zhang et al 2016 and Bornholdt et al 2016 as recited supra teaches monoclonal antibodies that strongly binds to sGP of Ebola virus and is a functional equivalent alternative for the claimed nanobody sGP7 and sGP49 to develop the claimed methods and the inventions. It would have been obvious to one of the ordinary skills to modify the prior art teachings of Gehrke et al incorporating the mAbs disclosed by Zhang et al 2016, and Bornholdt et al 2016 to replace nanobodies of Liu et al and arrive at the method of claim 1.
There would have been a reasonable expectation of success to arrive at the invention of instant claim 1 based on the applied prior art teachings as recited supra. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), See, MPEP § 2143, examples of rationales, A-G.
Claims 5-6: The combined prior art teachings of Gehrke et al, Liu et al (nanobodies binding to sGP), Zhang et al 2016, and Bornholdt et al 2016 (mAbs binding to sGPs ) and additional prior arts applied as recited supra teaches the method of claim 1 and is incorporated here by reference in entirety.
Claim 5: Gehrke et al further teaches the instant claim 5 limitation, the method of claim 1, wherein the detection step comprises determining a change in absorbance at a resonance wavelength of the AuNPs and/or the other MNPs. Gehrke et al discloses a UV-vis absorption spectra of plain gold nanostars (blue dashed), NStar conjugated to antibodies (blue solid line) and red NSs conjugated to antibodies (red solid line) indicating change in absorbance at a resonance wavelength of the AuNPs and/or the other MNPs (See, paragraph [0019], Figure 2-E, and associated legend). The inventions of Gehrke et al comprise an Ebola virus glycoprotein antigen detection in a sample using antibodies conjugated to a gold/silver or metallic nanoparticle.
Claim 6: Gehrke et al further teaches instant claim 6 limitation, the method of claim 1, comprising quantifying an amount of the glycoproteins and/or the Ebola virus in the sample. Gehrke et al discloses quantification (ng/ml) of viral antigen (NS1 of Dengue virus) in the same prior art disclosing Ebola virus glycoprotein antigen detection (See, paragraphs [144], [149], [173] and Figures 4B-4I). The inventions of Gehrke et al comprise an Ebola virus glycoprotein antigen detection in a sample using antibodies conjugated to a gold/silver or metallic nanoparticle.
It would have been obvious to one of the ordinary skills in the art before the effective filing date of the claimed invention to modify the combined prior art teachings as applied to the method of claim 1 as recited supra with additional teachings of Gehrke et al on the added claim 5 and 6 limitation as recited supra to arrive at the inventions of claims 5-6. The motivation would have been to develop a measurable diagnostic outcome by using change in absorbance at a resonance wavelength of AuNP or other MNPs to determine positive and negative measurement cutoff for content of sGP or Ebola virus in a sample and to quantify the sGP antigen or Ebola virus. One of the ordinary skills in the art would have a reasonable expectation of success to arrive at the invention of claims 5 and claim 6 given the applied prior art teachings as recited supra. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), See, MPEP § 2143, examples of rationales, A-G.
15. Claim 7 is rejected under 35 U.S.C. 103 as being unpatentable over combined teachings of Gehrke et al 2018 (US20180372755A1 published 27 December 2018), Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971), Liu et al 2017 (published in Microb Cell Fact (2017) 16:223), Ning et al 2017 (Virologica Sinica, 2017, 32 (1), p. 3–15), Zhang et al 2016 (Sci Rep 6, 25856, p. 1-15), Bornholdt et al 2016 (Science. 2016 Mar 4;351(6277):1078-83), Szunerits et al 2020 (Nanomaterials (Basel). 2020 Jun 29;10(7):1271), Wu et al 2020 (ACS Applied Nano Materials, 3(10), 9560-9580), Hung et al 2014 (Nanomedicine. 2014 May;10(4):819-29), Alejo-CanchoIet al 2020 (PLoS Negl Trop Dis. 2020 Feb 18;14(2)), Koh et al 2020 (Sensors 2009, 9, 8130-8145) as applied to claim 1, 3 and 5-6 above and further in view of Henson et al. 2017 (published in Material Matters, 2017, Vol 12-1).
Claim 7: The combined prior art teachings as applied as recited supra teaches the method of claim 1 and is incorporated here by reference in entirety, however, do not teach the added limitation of claim 7.
Henson et al is in the field of gold nanoparticles and teaches instant claim 7 limitation (the method of claim 1), further comprising centrifuging the aggregations of the bound glycoproteins prior to and/or during the detecting step. Henson et al. discloses centrifugation results pelleting and thus aggregation of gold nanoparticles (see page 29, column 1, section on insufficient purification; page 27 figure 2 and legends).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the prior art teaching on method of claim 1 to incorporate the teachings of Henson et al on-gold nanoparticle aggregate pellet formation by centrifugation to arrive at the invention of claim 7. The motivation would have been to pellet the heavier aggregates of the complex formed by Ebolavirus glycoprotein-antibody-conjugated with gold nanoparticles to obtain concentrated preparation for use in detection step to increase the sensitivity of the diagnostic assay. One of the ordinary skills in the art would have a reasonable expectation of success to arrive at the invention of claim 7 given the applied prior art teachings as recited supra. It is similar to some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the claimed invention. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), examples of rationales, G.
16. Claim 8 is rejected under 35 U.S.C. 103 as being unpatentable over combined teachings of Gehrke et al 2018 (US20180372755A1 published 27 December 2018), Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971), Liu et al 2017 (published in Microb Cell Fact (2017) 16:223), Ning et al 2017 (Virologica Sinica, 2017, 32 (1), p. 3–15), Zhang et al 2016 (Sci Rep 6, 25856, p. 1-15), Bornholdt et al 2016 (Science. 2016 Mar 4;351(6277):1078-83), Szunerits et al 2020 (Nanomaterials (Basel). 2020 Jun 29;10(7):1271), Wu et al 2020 (ACS Applied Nano Materials, 3(10), 9560-9580), Hung et al 2014 (Nanomedicine. 2014 May;10(4):819-29), Alejo-CanchoIet al 2020 (PLoS Negl Trop Dis. 2020 Feb 18;14(2)), Koh et al 2020 (Sensors 2009, 9, 8130-8145) as applied to claim 1, 3 and 5-6 above, and further in view of Albert et al. 2009 (published in Nanotechnology, 2009, Vol 20).
Claim 8: The combined prior art teachings as applied as recited supra teaches the method of claim 1 and is incorporated here by reference in entirety, however, do not teach the added limitation of claim 8.
Albert et al is in the field of gold nanoparticles and teaches the instant claim 8 limitation, (the method of claim 1), further comprising freezing the aggregations of the bound glycoproteins prior to the detecting step. Albert et al disclose freezing results in aggregation of gold-nanoparticles at the bottom of cuvettes; the TEM images revealing increased coalescence of AuNRs exposed to −20 ◦C for up to 6 h. (See, page 4, figure 3-B and figure 4 with associated legends; page 5, figure-5 with associated legends).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the prior art teaching on method of claim 1 to incorporate the teachings of Albert et al on-gold nanoparticle increased coalescence formation by freezing to arrive at the invention of claim 8. The motivation would have been to form increased coalescence of the complex formed by Ebolavirus glycoprotein-antibody-conjugated with gold nanoparticles to obtain concentrated preparation for use in detection step to increase the sensitivity of the diagnostic assay. One of the ordinary skills in the art would have a reasonable expectation of success to arrive at the invention of claim 7 given the applied prior art teachings as recited supra. It is similar to some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the claimed invention. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), examples of rationales, G.
17. Claim 9 is rejected under 35 U.S.C. 103 as being unpatentable over combined teachings of Gehrke et al 2018 (US20180372755A1 published 27 December 2018), Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971), Liu et al 2017 (published in Microb Cell Fact (2017) 16:223), Ning et al 2017 (Virologica Sinica, 2017, 32 (1), p. 3–15), Zhang et al 2016 (Sci Rep 6, 25856, p. 1-15), Bornholdt et al 2016 (Science. 2016 Mar 4;351(6277):1078-83), Szunerits et al 2020 (Nanomaterials (Basel). 2020 Jun 29;10(7):1271), Wu et al 2020 (ACS Applied Nano Materials, 3(10), 9560-9580), Hung et al 2014 (Nanomedicine. 2014 May;10(4):819-29), Alejo-CanchoIet al 2020 (PLoS Negl Trop Dis. 2020 Feb 18;14(2)), Koh et al 2020 (Sensors 2009, 9, 8130-8145) as applied to claim 1, 3 and 5-6 above, and further in view of Le et al. 2018 (published in Journal of Nanoscience and Nanotechnology, 2018, Vol 18, 1083-1088).
Claim 9: The combined prior art teachings as applied as recited supra teaches the method of claim 1 and is incorporated here by reference in entirety, however, do not teach the added limitation of claim 9.
Le et al is the field of virus and gold nanoparticles based viral antigen detection assay development. Le et al teaches the added limitation of claim 9, comprising drop casting the aggregations of the bound glycoproteins prior to the detecting step. Le et al teaches immunoreaction-mediated aggregation of gold nanoparticles for sensitive and selective assay of Hepatitis B surface antigen and demonstrates utility of Transmission Electron Microscopy (TEM) and dynamic light scattering analysis wherein the sample films for TEM analysis were formed by dropping (drop casting) the diluted solutions of AuNPs suspension on a holey carbon mesh grids (400 meshs) and air dried at room temperature and further dynamic light scattering analysis for detection step (See, abstract, page 1084-1085, section 2.4. Transmission Electron Microscopy, Figure 2) and sensitive detection of the target viral antigen in the range of 0.5 ng/ml to 50 ng/ml (See, page 1086, figure 4).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the prior art teachings on method of claim 1 as recited supra to incorporate the teachings of Le et al on the drop casting immunocomplex of HBV surface antigen bound to the gold nanoparticles conjugated antibody and achieve the instant claim 9 invention by drop casting the aggregations of the bound glycoproteins prior to the detecting step. The motivation would have been to form the sample films for TEM analysis by dropping (drop casting) the aggregated immunocomplexes of Ebolavirus glycoprotein- bound to gold nanoparticle conjugated antibodies on a holey carbon mesh grids (400 meshs) and air dried at room temperature and further perform dynamic light scattering analysis for detection step and obtain higher sensitive detection of the target viral antigen in the range of 0.5 ng/ml to 50 ng/ml (See, Le aet al; abstract and figure 4). One of the ordinary skills in the art would have a reasonable expectation of success to arrive at the invention of claim 7 given the applied prior art teachings as recited supra. It is similar to combining prior art elements according to known methods to yield predictable results. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), examples of rationales, A.
18. Claims 12 and 14 are rejected under 35 U.S.C. 103 as being unpatentable over combined teachings of Gehrke et al 2018 (US20180372755A1 published 27 December 2018), Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971), Liu et al 2017 (published in Microb Cell Fact (2017) 16:223), Ning et al 2017 (Virologica Sinica, 2017, 32 (1), p. 3–15), Zhang et al 2016 (Sci Rep 6, 25856, p. 1-15), Bornholdt et al 2016 (Science. 2016 Mar 4;351(6277):1078-83), Szunerits et al 2020 (Nanomaterials (Basel). 2020 Jun 29;10(7):1271), Wu et al 2020 (ACS Applied Nano Materials, 3(10), 9560-9580), Hung et al 2014 (Nanomedicine. 2014 May;10(4):819-29), Alejo-CanchoIet al 2020 (PLoS Negl Trop Dis. 2020 Feb 18;14(2)), Koh et al 2020 (Sensors 2009, 9, 8130-8145) as applied to claims 1, 3, 5-6 and 10 above.
Claims 12 and 14: The combined prior art teachings as applied as recited supra teaches the method of claim 10 and is incorporated here by reference in entirety.
Regarding claim 12 added limitation Gehrke et al further discloses paper-based rapid diagnostic tests (RDTs) are convenient, robust, and can be read out within minutes and the inventions of Gehrke et al comprises an Ebola virus glycoprotein antigen detection in samples using antibodies conjugated to a gold/silver or metallic nanoparticle (See, abstract, paragraph [0003], [0007] and [0131]).
Regarding claim 14 added limitation Gehrke et al discloses added limitation of claim 14, the method of claim 10, wherein the sample comprises blood, plasma, serum, saliva, sputum, or urine. Gehrke et al discloses a sample of biological origin, or a sample derived from the sample of biological origin, preferably from human patient. The biological samples include, but are not limited to, blood, plasma, serum, saliva, cerebral spinal fluid, ……, sputum, urine, stool, tear, ….., respiratory, intestinal, or genitourinary tract, organ, cell culture, cell culture constituent, tissue sample, tissue section, whole cell, cell constituent, or cell smear (See, paragraph [0110]).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the prior art teaching on method of claim 10 to incorporate the additional teachings of Gehrke et al on convenient, robust, and can be read out within minutes for presence of Ebola virus sGP or Ebola virus in variety of available biological samples as recited supra to arrive at the invention of claims 12 and 14. The motivation would have been to develop a rapid high throughput Ebola antigen diagnostic assay for emergency treatment of the subjects diagnosed of Ebola virus infection. One of the ordinary skills in the art would have a reasonable expectation of success to arrive at the invention of claims 12 and 14 given the applied prior art teachings as recited supra. It is similar to some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the claimed invention. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), examples of rationales, G.
19. Claims 11 and 13 are rejected under 35 U.S.C. 103 as being unpatentable over combined teachings of Gehrke et al 2018 (US20180372755A1 published 27 December 2018), Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971), Liu et al 2017 (published in Microb Cell Fact (2017) 16:223), Ning et al 2017 (Virologica Sinica, 2017, 32 (1), p. 3–15), Zhang et al 2016 (Sci Rep 6, 25856, p. 1-15), Bornholdt et al 2016 (Science. 2016 Mar 4;351(6277):1078-83), Szunerits et al 2020 (Nanomaterials (Basel). 2020 Jun 29;10(7):1271), Wu et al 2020 (ACS Applied Nano Materials, 3(10), 9560-9580), Hung et al 2014 (Nanomedicine. 2014 May;10(4):819-29), Alejo-CanchoIet al 2020 (PLoS Negl Trop Dis. 2020 Feb 18;14(2)), Koh et al 2020 (Sensors 2009, 9, 8130-8145) as applied to claims 1, 3, 5-6 and 10, and further in view of Reynard and Volchkov 2019 (US10266584B2 patent granted on 23 April 2019) and Nouvellet et al 2015 (published in Nature (2015) 528, S109–S116).
Claims 11 and 13: The combined prior art teachings as applied as recited supra teaches the method of claim 10 and is incorporated here by reference in entirety. However, does not teach the added limitations of the claim 11, comprising administering one or more therapies to the subject when the Ebola virus is detected in the sample (claim 11 limitation); and, does not explicitly recite added limitation of the claim 13 comprising repeating the method using one or more longitudinal samples obtained from the subject (claim 13 limitation).
Reynard and Volchkov disclose administering both prophylactic or preventive treatment as well as curative or disease modifying treatment, for Ebola virus infection using antibodies or drugs, including treatment of subjects at risk of contracting the disease or suspected to have contracted the disease as well as subjects who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse. The antibody is used in combination with at least one antiviral compound; the anti-viral compound is selected from polymerase inhibitors. The entire invention is directed to antibodies specific to glycoprotein (GP) of Ebolavirus and uses for the treatment and diagnosis of ebola virus infection (See, US10266584B2, title and abstract, column 22 lines 1-33 and 23 lines 1-65).
Nouvellet et al teaches the role or use of rapid Ebola virus antigen detection diagnostics test in managing Ebola epidemics that are infected, exposed and negative patients (See, page S109- abstract; figure 1 and associated legends on page S111).
It would have been obvious to one of ordinary skill in the art, at the time of the claimed invention, to modify the combined prior arts teachings method claim 10 on obtaining the samples from the subject as disclosed by Gehrke et al to incorporate the teachings of Nouvellet et al on use of rapid Ebola virus antigen detection diagnostics test in managing Ebola epidemics and use the approach to monitor the effectiveness of Ebola virus therapy administered in the instant claim 11 as taught by Reynard and Volchkov. The motivation for repeating the rapid diagnostic method using one or more longitudinal samples obtained from the subject receiving anti-Ebola virus therapy would provide information on effectiveness of the treatment and manage the patient health. One of the ordinary skills in the art would have a reasonable expectation of success to arrive at the invention of claims 11 and 13 given the applied prior art teachings as recited supra. It is similar to some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the claimed invention. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), examples of rationales, G.
20. Claims 15-17 are rejected under 35 U.S.C. 103 as being unpatentable over combined teachings of Gehrke et al 2018 (US20180372755A1 published 27 December 2018), Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971), Liu et al 2017 (published in Microb Cell Fact (2017) 16:223), Ning et al 2017 (Virologica Sinica, 2017, 32 (1), p. 3–15), Zhang et al 2016 (Sci Rep 6, 25856, p. 1-15), Bornholdt et al 2016 (Science. 2016 Mar 4;351(6277):1078-83), Szunerits et al 2020 (Nanomaterials (Basel). 2020 Jun 29;10(7):1271), Wu et al 2020 (ACS Applied Nano Materials, 3(10), 9560-9580), Hung et al 2014 (Nanomedicine. 2014 May;10(4):819-29), Alejo-CanchoIet al 2020 (PLoS Negl Trop Dis. 2020 Feb 18;14(2)), Koh et al 2020 (Sensors 2009, 9, 8130-8145) as applied to claims 1, 3, 5-6 and 10, and further in view of Rica and Stevens et al. 2013 (published in Nat Protoc, 8, 1759–1764).
Claims 15-17: The combined prior art teachings as applied as recited supra teaches the method of claim 1 and is incorporated here by reference in entirety.
Claim 15: The method of claim 1, wherein the detecting step comprises measuring a colorimetric change when the one or more aggregations of the bound sGPs form with one another.
Regarding Claim 15: Gehrke et al further teaches added limitation of claim 15, wherein the detecting step comprises measuring a colorimetric change when the one or more aggregations of the bound glycoproteins form with one another. Gehrke et al discloses colorimetric differential detection of Ebola virus glycoprotein using antibodies labelled with unique colorimetric label is selected from, gold nanoparticles, colored latex beads, dye labeled beads, silver nanoparticles, quantum dots, up converting phosphors, and organic fluorophores, with unique spectral emission (See, paragraph [0008-0011, 0108-0109, 0116, 0125-0127, 0143, 0166], claim 1,3-6,11,15, and 22).
Rica and Stevens et al is in the diagnostic virology art and teaches a plasmonic colorimetric ELISA for detection of HIV p24 antigen or prostate-specific antigen (PSA) by capture monoclonal antibodies, primary antibodies (polyclonal antibodies) specific to respective p24 or PSA target antigens (See, Rica and Stevens et al, pages 1759-1764, title, abstract, page 1760m figure 1, and see entire article). Rica and Stevens et al teaches instant claim 15 limitation, an approach for quantifying the concentration of analyte (HIV p24 or PSA antigen) by using recently reported plasmonic nano-sensors that work with inverse sensitivity for those applications in which the quantification of the analyte at ultralow concentration is required; the method is also based on the enzyme-guided growth of gold nanoparticles, allows for a similar limit of detection as plasmonic ELISA and has a dynamic range spanning over four or five orders of magnitude depending on the sample matrix (See , page 1760, section on comparison with other methods). It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the method of claim 1 to incorporate the teachings of Rica and Stevens et al on plasmonic colorimetric ELISA used for detection of HIV p24 antigen or prostate-specific antigen (PSA) and modify the assay for Ebola virus glycoprotein detection using specific capture monoclonal antibodies and primary antibodies with gold or plasmonic metallic nanoparticles as per teachings of Rica and Stevens et al to arrive at the invention of instant claim 15. The motivation would have been to develop a sensitive assay to detect an Ebola virus secreted glycoprotein in samples from subjects. One of the ordinary skills in the art would have a reasonable expectation of success to arrive at the invention of claim 15 given the applied prior art teachings as recited supra. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), See, MPEP § 2143, examples of rationales, A-G.
Claim 16: The method of claim 15, comprising visually detecting the colorimetric change when the one or more aggregations of the bound sGP form with one another.
Rica and Stevens et al is in the diagnostic virology art and teaches instant claim 16 limitation visually detecting the colorimetric change when one or more aggregations of the bound antigen form with one another using an example of HIV p24 or PSA antigen (See, page 1760, figure 1-c, 1764, figure 3, methods, entire article) and demonstrates visual detection using plasmonic ELISA showing blue color. It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the method of claim 15 to incorporate the teachings of Rica and Stevens et al on visual detection of plasmonic colorimetric ELISA used for detection of HIV p24 antigen or prostate-specific antigen (PSA) and modify the assay for Ebola virus glycoprotein detection using specific capture monoclonal antibodies and primary antibodies with gold or plasmonic metallic nanoparticles as per teachings of Rica and Stevens et al to arrive at the invention of instant claim 16. The motivation would have been to develop a visual qualitative colorimetric assay to detect an Ebola virus/ Ebola virus secreted glycoprotein in samples from subjects to obviate a need for reading device (See, Rica and Stevens et al). One of the ordinary skills in the art would have a reasonable expectation of success to arrive at the invention of claim 16 given the applied prior art teachings as recited supra. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), See, MPEP § 2143, examples of rationales, A-G.
Claim 17: The method of claim 15, comprising visually detecting the colorimetric change when the one or more aggregations of the bound sGPs form with one another.
Regarding claim 17: Rica and Stevens et al teaches instant claim 17 limitation detecting the colorimetric change when one or more aggregations of the bound antigen form with one another using an example of HIV p24 or PSA antigen (See, page 1764, figure 3, methods, entire article) and demonstrates detection using plasmonic ELISA showing blue color at different concentration of target antigens. It would have been obvious to one of ordinary skill in the art, at the time of the claimed invention, to modify the method of detection of Ebola virus glycoprotein using gold or plasmonic metallic nanoparticle conjugated antibodies of claim 1 as disclosed by Gehrke et al to incorporate the teachings of Rica and Stevens et al on detection of plasmonic colorimetric ELISA used for detection of HIV p24 antigen or prostate-specific antigen (PSA) and modify the assay for Ebola virus glycoprotein detection using specific capture monoclonal antibodies and primary antibodies with gold or plasmonic metallic nanoparticles as per teachings of Rica and Stevens et al to arrive at the invention of instant claim 17. The motivation would have been to develop a visual qualitative colorimetric assay to detect an Ebola virus/ Ebola virus secreted glycoprotein in samples from subjects to obviate a need for reading device (See, Rica and Stevens et al). One of the ordinary skills in the art would have a reasonable expectation of success to arrive at the invention of claim 17 given the applied prior art teachings as recited supra. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), See, MPEP § 2143, examples of rationales, A-G.
21. Claims 18 is rejected under 35 U.S.C. 103 as being unpatentable over Gehrke et al 2018 (US20180372755A1 published 27 December 2018), and further in view of Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971), Liu et al 2017 (published in Microb Cell Fact (2017) 16:223), Ning et al 2017 (Virologica Sinica, 2017, 32 (1), p. 3–15), Zhang et al 2016 (Sci Rep 6, 25856, p. 1-15), Bornholdt et al 2016 (Science. 2016 Mar 4;351(6277):1078-83), Szunerits et al 2020 (Nanomaterials (Basel). 2020 Jun 29;10(7):1271), Wu et al 2020 (ACS Applied Nano Materials, 3(10), 9560-9580), Hung et al 2014 (Nanomedicine. 2014 May;10(4):819-29), Alejo-CanchoIet al 2020 (PLoS Negl Trop Dis. 2020 Feb 18;14(2)), Koh et al 2020 (Sensors 2009, 9, 8130-8145).
Claim 18: Gehrke et al further discloses a device, a rectangular lateral flow assay device (LFA device) for point of care (See, abstract and figure 1) to detect an Ebola virus or Ebola virus GP1 antigen in a biological sample obtained from a subject/human patient infected with Ebola virus. The LFA devices comprise a porous matrix, nitrocellulose, that absorb a liquid sample and promotes capillary action along the nitrocellulose. The device has a sample loading chamber that comprise nitrocellulose sample pad, connected by a conjugate pad that comprise detecting antibodies conjugated to gold or other plasmonic metal nanoparticles and joined by a capture-detection area/chamber of nitrocellulose along the central region comprising immobilized capture antibodies to specifically capture Ebola virus/ Ebola virus glycoproteins (See, abstract, claims 1-14, paragraph [0008], [0009], [0099], [0101], [0110], [0111]); the sample is added drop-wise in a sample loading chamber on one end on the LFA device (See, Figure 1, paragraph [0104], [0105]); the sample migrates through the conjugate pad comprising gold nanoparticle conjugated detector antibody and forms a glycoprotein- gold nanoparticle conjugated detector antibody complex, the complex moves to the antigen capture chamber of nitrocellulose membrane and in a positive sample forms a capture antibody-glycoprotein-gold nanoparticle conjugated detector antibody complex, becomes immobilized and aggregated immunocomplexes produces a distinct signal, for example a colored line in the capture-detection area/chamber of nitrocellulose. The LFA has a negative, positive control and text line, and some LFA may have more than one test line for multiplex testing for multiple infectious disease biomarkers and are one example of a “multiplexed immunoassay” (See, figure 1, paragraphs [0008], [0009], [0104], [0105]).
The combined prior art teachings, obviousness analysis and motivation as applied and recited supra rendering obvious the method of claim 1 is incorporated here by reference in entirety as applicable to claim 18 limitations.
Chen et al 2022 as recited supra under 35 U.S.C. 102(a)(1) rejection teaches sGP7 and sGP49 nanobody CDR1, CDR2 and CDR3 amino acid sequences SEQ ID NO: 1-3 (sGP7 binding nanobody), and SEQ ID NO: 4-6 (sGP49 binding nanobody) as recited supra with 100% amino acid identity and are incorporated here in entirety.
Hung et al 2014 is in the virology diagnostic art and teaches magnetic nanoparticle-based immunoassay for rapid detection of influenza infections by using an integrated microfluidic system (See, abstract, Fig 1-6, entire article).
Alejo-Cancho et al 2020 is in the virology diagnostic art and teaches evaluation of a novel microfluidic immunomagnetic agglutination assay method for detection of dengue virus NS1 antigen (See, entire article, Fig 2).
Koh et al 2020 is in the viral diagnostic art and reviews magnetic nanoparticle sensors (See, abstract and entire review).
Additionally, claim 18 is directed to a device, without recitation of structure, but merely contain the assay components recited in claim 1. In this regard, consistent with the specification disclosure, conventional apparatuses comprising “reaction chambers” and systems further containing means of detecting antibody (nanobody) antigen binding were known in the prior art. See e.g. Ye et al. March 2020: 10(10);4359-4373 and prior art cited therein.
It would have been obvious to one of the ordinary skill in the art before the effective filing date of the claimed invention to modify the prior art teachings of Gehrke et al as recited supra to incorporate the teachings of Chen et al 2022, Liu et al comprising the nanobodies binding to heavily glycosylated glycoprotein of Ebola virus (representing sGP) or functional equivalent mAbs binding to sGP as taught by Zhang et al 2016 and Bornholdt et al 2016 as recited supra as an alternative for the claimed nanobody sGP7 and sGP49 (See, claim 1 rejection, supra), teachings of Alejo-Cancho et al, Hung et al, Koh et al on a device and system to arrive at the invention of claim 18. The motivation would have been to design and develop a device for rapid detection and quantification of Ebola virus sGP using the nanobodies in a clinical sample from subject or experimental samples or standard purified sGP antigen for treatment of the subject and for commercial advantage. There would have been a reasonable expectation of success to arrive at the invention of instant claim 18 based on the applied prior art teachings as recited supra. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), See, MPEP § 2143, examples of rationales, A-G.
22. Claims 19-20 are rejected under 35 U.S.C. 103 as being unpatentable over Gehrke et al 2018 (US20180372755A1 published 27 December 2018), and further in view of Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971), Liu et al 2017 (published in Microb Cell Fact (2017) 16:223), Ning et al 2017 (Virologica Sinica, 2017, 32 (1), p. 3–15), Zhang et al 2016 (Sci Rep 6, 25856, p. 1-15), Bornholdt et al 2016 (Science. 2016 Mar 4;351(6277):1078-83), Szunerits et al 2020 (Nanomaterials (Basel). 2020 Jun 29;10(7):1271), Wu et al 2020 (ACS Applied Nano Materials, 3(10), 9560-9580), Hung et al 2014 (Nanomedicine. 2014 May;10(4):819-29), Alejo-CanchoIet al 2020 (PLoS Negl Trop Dis. 2020 Feb 18;14(2)), Koh et al 2020 (Sensors 2009, 9, 8130-8145), Qu et al. 2020 (published on 13 January 2020 in Nanoscale Research Letters (2020) 15:10) and Ye et al 2020 (Published on 15 March 2020 in Theranostics 2020, Vol. 10, Issue 10).
Claim 19: Gehrke et al teaches the instant claim 19 limitation, a device, a rectangular lateral flow assay device (LFA device) for point of care (See, abstract and figure 1) to detect an Ebola virus or Ebola virus glycoprotein 1 (GP1) antigen in a biological sample obtained from a subject/human patient infected with Ebola virus. The LFA devices comprise a porous matrix, nitrocellulose, that absorb a liquid sample and promotes capillary action along the nitrocellulose. The device has a sample loading chamber that comprise nitrocellulose sample pad, connected by a conjugate pad that comprise detecting antibodies conjugated to gold or other plasmonic metal nanoparticles and joined by a capture-detection area/chamber of nitrocellulose along the central region comprising immobilized capture antibodies to specifically capture Ebola virus/ Ebola virus glycoproteins by binding to antigenic sites/epitopes (See, abstract, claims 1-14, paragraph [0008], [0009], [0099], [0101], [0110], [0111]); the sample is added drop-wise in a sample loading chamber on one end on the LFA device (See, Figure 1, paragraph [0104], [0105]); the sample migrates through the conjugate pad comprising gold nanoparticle conjugated detector antibody and forms a glycoprotein- gold nanoparticle conjugated detector antibody complex, the complex moves to the antigen capture chamber of nitrocellulose membrane and in a positive sample forms a capture antibody-glycoprotein-gold nanoparticle conjugated detector antibody complex is formed by binding of capture antibodies to antigenic sites/epitopes of glycoprotein, becomes immobilized and aggregated immunocomplexes produces a distinct signal, for example a colored line in the capture-detection area/chamber of nitrocellulose. The LFA has a negative, positive control and text line, and some LFA may have more than one test line for multiplex testing for multiple infectious disease biomarkers and are one example of a “multiplexed immunoassay” (See, figure 1, paragraphs [0008], [0009], [0104], [0105]). The LFA assay allows diagnosis at point of care (POC) within minutes and readout by eye or mobile phone (See abstract, [0006]); for readout, …. image analysis to enable diagnosis simply by distinguishing the color of the assay (See, [0119]); the approach of analyzing the RGB signal of the test line could be used to detect if an emerging biomarker is present in the sample that is not the one the antibodies are screened for. This approach could also be applied to test lines that use fluorescent or colorimetric beads of different colors for readout (See para [0166]).
The combined prior art teachings, obviousness analysis and motivation as applied and recited supra rendering obvious the method of claim 1 is incorporated here by reference in entirety as applicable to claim 19 limitations. The teachings of Liu et al comprising the nanobodies binding to heavily glycosylated glycoprotein of Ebola virus (representing sGP) or functional equivalent mAbs binding to sGP as taught by Zhang et al 2016 and Bornholdt et al 2016 as recited supra as an alternative for the claimed nanobody sGP7 and sGP49 (See, claim 1 rejection, supra), teachings of Alejo-Cancho et al, Hung et al, Koh et al on a device and system.
Chen et al 2022 as recited supra under 35 U.S.C. 102(a)(1) rejection teaches sGP7 and sGP49 nanobody CDR1, CDR2 and CDR3 amino acid sequences SEQ ID NO: 1-3 (sGP7 binding nanobody), and SEQ ID NO: 4-6 (sGP49 binding nanobody) as recited supra with 100% amino acid identity and are incorporated here in entirety.
Hung et al 2014 is in the virology diagnostic art and teaches magnetic nanoparticle-based immunoassay for rapid detection of influenza infections by using an integrated microfluidic system (See, abstract, Fig 1-6, entire article).
Alejo-Cancho et al 2020 is in the virology diagnostic art and teaches evaluation of a novel microfluidic immunomagnetic agglutination assay method for detection of dengue virus NS1 antigen (See, entire article, Fig 2).
Koh et al 2020 is in the viral diagnostic art and reviews magnetic nanoparticle sensors (See, abstract and entire review).
Gehrke et al does not teach the claim 19 limitation, an electromagnetic radiation detection apparatus positioned, or positionable, within sufficient proximity to the device to detect one or more colorimetric changes produced in or on the reaction chamber or substrate when one or more aggregations of the bound glycoproteins form with one another in or on the reaction chamber or substrate.
Qu et al teaches the lateral flow assay for immunochromatographic detection and quantification of human chorionic gonadotropin (HCG) biomarker using plasmonic gold nanoparticle conjugated specific antibody and use of light sources located at the near infrared (NIR) region of the electromagnetic spectrum allows heat generation using the plasmonic properties of anisotropic gold nanoparticles to prevent the absorption of the light by the majority of the molecules from a biological origin, particularly blood. The laser diode light source of the detection apparatus is positioned in close proximity of the reaction chamber to allow detection and quantification. Qu et al optimized the operating conditions for the device using conduction and radiation thermal sensing modes allowing the quantification of LFA. The limit of detection of the strips merely containing nanoparticles was decreased by 5-fold (conduction mode) and 12-fold (radiation mode) compared to traditional visual detection. The effect of the ambient temperature was studied for both methods of detection showing that the radiation mode was more affected by the ambient temperature than the conduction mode. To validate the thermal sensing method, human chorionic gonadotropin (HCG) biomarker was quantified using our LFA strips, obtaining a detection limit of 2.8 mIU/mL when using the radiation method of detection. (See, abstract; page 2, scheme 1; column 2 paragraph 2; page 4, figure 1; page 7 figure 3; pages 8-9, 11 figures 4-6; see entire article).
Ye et al in a review teaches signal amplification and quantification on lateral flow assays by laser excitation of plasmonic nanomaterials using gold nanoparticles Raman molecules and antibody-coated hollow GNPs as Surface-enhanced Raman scattering (SERS) tags for the LFA; positioning detection apparatus in proximity of detection chamber; gold nanoparticles (GNPs) can be excited by the laser and convert the absorbed energy into enhanced electromagnetic field or heat due to the LSPR. Such responses of GNPs provide new sensing modes that have been successfully integrated on LFA, (See, figure 2A, figure 3; page 4369- summary outlook; see entire article).
Additionally, claim 19 is directed to a device and a system, without recitation of structure, but merely contain the assay components recited in claim 1. In this regard, consistent with the specification disclosure, conventional apparatuses comprising “reaction chambers” and systems further containing means of detecting antibody (nanobody) antigen binding were known in the prior art. See e.g. Ye et al. March 2020: 10(10);4359-4373 and prior art cited therein.
It would have been obvious to one of the ordinary skills in the art before the effective filing date of the claimed invention to modify the prior art teachings of Gehrke et al to incorporate the combined prior art teachings of Liu et al on Ebola virus or GP1, sGP binding nanobodies or mAbs (functional equivalent) binding to sGP as taught by Zhang et al 2016 and Bornholdt et al 2016 as recited supra as an alternative and conjugate to gold nanoparticles for plasmonic colorimetric detection and incorporate teachings of Qu et al, Le et al on electromagnetic radiation detection apparatus to arrive at the invention or the system of the claim 19. The motivation would have been to enhance sensitivity of the assay and to reduce the noise by preventing absorption of the light by the majority of the molecules from a biological origin, particularly blood (See, Qu et al); Surface-enhanced Raman scattering (SERS)- based LFA with enhanced detection sensitivity (See, Ye aet al-figure 2 legend). There would have been a reasonable expectation of success to arrive at the invention of instant claim 19 based on the applied prior art teachings as recited supra. It is similar to combining prior art elements according to known methods to yield predictable results. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), examples of rationales, A.
Claim 20: The combined prior art teachings as applied to claim 19 and recited supra teaches the invention of claim 19; however, does not teach the added limitations of claim 20.
Qu et al and Ye et al further teaches the added limitation of instant claim 20.
Ye et al teaches the electromagnetic radiation detection apparatus comprises a spectrometer as taught by figure 2 schematic representation of a portable SERS reader with line-focused optical fiber probe showing a Laser light diode probe in close proximity of the detection chamber of the lateral flow assay and attached spectrophotometer. As a proof-of-principle, bulky Raman microscope system was used to measure the SERS signal of LFA, yet impractical for POC deployment. Recently, …. reported a portable SERS reader designed for rapid scanning of the LFA strips. The compact setup mainly composed of an optical fiber probe and a 785 nm diode laser (figure C, page 4362, column 2, paragraph 1-section on Reader development); wherein the electromagnetic radiation detection apparatus comprises a light-emitting diode (LED) that transmits light into and/or through the reaction chamber or substrate and a photodetector that detects light from the reaction chamber or substrate is taught by Ye et al (See entire article and figures 1-6) as recited supra and Qu et al as shown in figure 1 uses a laser light diode positioned closely to the detection chamber of the lateral flow assay device (See, Qu et al, page 2, scheme 1 and figure 1-6, and entire article).
Additionally, claim 20 is directed to a device and a system, without recitation of structure, but merely contain the assay components recited in claim 1. In this regard, consistent with the specification disclosure, conventional apparatuses comprising “reaction chambers” and systems further containing means of detecting antibody (nanobody) antigen binding were known in the prior art. See e.g. Ye et al. March 2020: 10(10);4359-4373 and prior art cited therein.
It would have been obvious to one of the ordinary skills in the art, at the time of the claimed invention, to further modify the combined teachings as applied to claim 19 to incorporate additional teachings of Qu et al, Le et al on electromagnetic radiation detection apparatus comprising light emitting diode to arrive at the invention or the system of the claim 20. The motivation would have been to enhance sensitivity of the assay and to reduce the noise by preventing absorption of the light by the majority of the molecules from a biological origin such as blood (See, Qu et al); Surface-enhanced Raman scattering (SERS)- based LFA with enhanced detection sensitivity (See, Ye aet al-figure 2 legend).
There would have been a reasonable expectation of success to arrive at the invention of instant claim 20 based on the applied prior art teachings as recited supra. It is similar to combining prior art elements according to known methods to yield predictable results. See, KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007), examples of rationales, A.
Response to Applicant’s Arguments
Applicant’s arguments filed on 11/21/2025 with respect to claims 1, 3, and 5-20 have been considered but are moot because the new matter and new ground of rejection does not rely on only the prior applied reference but also include new references as prior arts of record for any teaching or matter specifically challenged in the argument.
Applicant’s argument: Rejections Under 35 U.S.C. § 103
Claims 1, 3, 5, 6, 10, 14 and 18 are rejected under 35 U.S.C. § 103 as allegedly being
obvious over Gehrke, and further in view of Liu, Ning, Zhang, Bornholdt, Szunerits, Wu, Hung, Alejo-Canchol and Koh. Claim 7 is rejected under 35 U.S.C. § 103 as allegedly being obvious over Gehrke, Liu, Ning, Zhang, Bornholdt, Szunerits, Wu, Hung, Alejo-Cancho, Koh as applied to claims 1, 3, 5 and 6 above and further in view of Henson et al. 2017 (Material Matters, 2017, Vol 12-1, hereinafter, "Henson"). Claim 8 is rejected under 35 U.S.C. § 103 as allegedly being obvious over Gehrke, Liu, Ning, Zhang, Bornholdt, Szunerits, Wu, Hung, Alejo-Cancho, Koh as applied to claims 1, 3, 5 and 6 above, and further in view of Albert et al. (Nanotechnology, 2009, Vol 20, hereinafter, "Albert"). Claim 9 is rejected under 35 U.S.C. § 103 as allegedly being obvious over Gehrke, Liu, Ning, Zhang, Bornholdt, Szunerits, Wu, Hung, Alejo-Cancho, Koh as applied to claims 1, 3, 5 and 6 above and further in view of Le et al. (Journal of Nanoscience and Nanotechnology, 2018, Vol 18, 1083-1088, hereinafter, "Le"). Claims 12 and 14 are rejected under 35 U.S.C. § 103 as allegedly being obvious over Gehrke, Liu, Ning, Zhang, Bornholdt, Szunerits, Wu, Hung, Alejo-Cancho, Koh as applied to claims 1, 3, 5, 6 and 10 above. Claims 11 and 13 are rejected under 35 U.S.C. § 103 as allegedly being obvious over Gehrke, Liu, Ning, Zhang, Bornholdt, Szunerits, Wu, Hung, Alejo-Cancho, Koh as applied to claims 1, 3, 5, 6 and 10, and further in view of Reynard et al. (U.S. Patent No. 10,266,584 B2, hereinafter, "Reynard") and Nouvellet et al. (Nature (2015) 528, S109-S116, hereinafter, "Nouvellet"). Claims 15-17 are rejected under 35 U.S.C. § 103 as allegedly being obvious over Gehrke, Liu, Ning, Zhang, Bornholdt, Szunerits, Wu, Hung, Alejo-Cancho, Koh as applied to claims 1, 3, 5, 6 and 10, and further in view of de la Rica et al. (Nat Protoc, 8, 1759-1764, hereinafter, "de la Rica"). In addition, claims 19 and 20 are rejected under 35 U.S.C. § 103 as allegedly being obvious over Gehrke, and further in view of Liu, Ning, Zhang, Bornholdt, Szunerits, Wu, Hung, Alejo- Canchol, Koh, Qu et al. (Nanoscale Research Letters (2020) 15:10, hereinafter, "Qu") and Ye et al. (Theranostics 2020, Vol. 10, Issue 10, hereinafter, "Ye").
As noted above, the Applicant has amended each of independent claims 1, 18, and 19, in relevant part, to further recite that complementarity-determining regions (CDRs) "of the sGP49 nanobody comprise the amino acid sequences of SEQ ID NOs:4-6" and those "of the sGP7 nanobody comprise the amino acid sequences of SEQ ID NOs:1-3". None of the cited art, considered individually or in any combination, teaches or suggests each and every element of claim 1, 18, or 19, particularly as amended to include the amino acid sequence information of the CDRs of the recited sGP49 and sGP7 nanobodies. As a consequence, the Office Action fails to establish a primafacie case of obviousness with respect to claim 1, 18, or 19 as amended on this exemplary basis alone. Moreover, the Office Action does not articulate any teaching, suggestion, or motivation in the cited art that would have led one of ordinary skill to modify that prior art to arrive at the claimed subject matter of any pending claim, particularly with any reasonable expectation of success at doing so. Accordingly, as claims 1, 18, and 19 as amended, and necessarily all of their dependent claims are non-obvious in view the cited art, the Applicant respectfully requests that this rejection be withdrawn.
In Response: Applicant’s arguments filed on 11/21/2025 with respect to claims 1, 3, and 5-20 rejection under obviousness have been considered but are not persuasive in view of new matter and new ground of rejection the arguments are moot, and the rejections are maintained.
Double Patenting (Modified)
23. The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
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The instant claims 1, 3, and 5-20 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 4-14, 18-20, 28-29, and 33-50 of copending Application No. 18247161 in view of Gehrke et al 2018 (US20180372755A1 published 27 December 2018), and further in view of Chen et al 2022 (published online on 01/06/2022 in Biosensors and Bioelectronics (2022) vol 202, 113971), Liu et al 2017 (published in Microb Cell Fact (2017) 16:223), Ning et al 2017 (Virologica Sinica, 2017, 32 (1), p. 3–15), Zhang et al 2016 (Sci Rep 6, 25856, p. 1-15), Bornholdt et al 2016 (Science. 2016 Mar 4;351(6277):1078-83), Szunerits et al 2020 (Nanomaterials (Basel). 2020 Jun 29;10(7):1271), Wu et al 2020 (ACS Applied Nano Materials, 3(10), 9560-9580), Hung et al 2014 (Nanomedicine. 2014 May;10(4):819-29), Alejo-CanchoIet al 2020 (PLoS Negl Trop Dis. 2020 Feb 18;14(2)), Koh et al 2020 (Sensors 2009, 9, 8130-8145) (additional all the prior art teachings applied to render obvious instant claims 1, 3, 5-20 as recited supra)
Both the instant claims 1, 3, and 5-20 and co-pending reference claims 1, 4-14, 18-20, 28-29, and 33-50 are similarly drawn to a method detecting an Ebola virus glycoprotein in a sample, inter alia, using antibodies or antigen binding portion thereof or nanobodies conjugated to plurality of gold nanoparticles (AuNPs) and/or other plasmonic metal nanoparticles (MNPs) and to detecting the bound Ebola virus secreted glycoprotein (sGP) - gold nanoparticle conjugated antibody aggregates in a lateral flow assay format with a readout by visual or machine. The instant claims 1, 3, and 5-20 are presented and recited similarly to the co-pending reference claims 1, 4-14, 18-20, 28-29, and 33-50.
The difference in the inventions claimed by the instant claim and co-pending claims is that instant claims are directed to detect an Ebola virus sGP detection using nanobody sGP7 and sGP49 that bind to first and second epitopes of sGP whereas copending claims are directed to detect secreted glycoprotein (sGP) by using one or more antibodies, or antigen binding portions thereof, or camelid nanobodies that bind to the epitope of a secreted glycoprotein (sGP) from the Ebola virus.
The combined prior art teachings rendering obvious claims 1, 3, and 5-20 and the obviousness analysis, motivations and expected reasonable success to arrive at the inventions of instant claims is incorporated here in entirety by reference to render obvious the reference application no. 18247161 claims 1, 4-14, 18-20, 28-29, and 33-50.
Therefore, before the effective filing date of the claimed invention, it would have been obvious to a person having ordinary skills in the art to have been motivated to modify the co-pending Application No. 18247161 reference claims 1, 4-14, 18-20, 28-29, and 33-50 with the incorporated teachings as recited supra.
Therefore, instant claims 1, 3, and 5-20 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over copending application no. 18247161 reference claims 1, 4-14, 18-20, 28-29, and 33-50 in view of the applied prior art teachings and obviousness analysis.
Such modifications, combining prior art elements, as recited, supra, according to known methods, would have had a reasonable expectation of success to have arrived at the claimed invention at the effective filing date of the instant application. This is analogous to some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the claimed invention in instant claims 1, 3, and 5-20. See KSR Int'l Co. v. Teleflex Inc., 550 U.S. 398, 415-421, 82 USPQ2d 1385, 1395-97 (2007) (see MPEP § 2143, example of rationales, G). For at least these reasons, instant claims 1, 3, and 5-20 are unpatentable over the co-pending reference Application No. 18247161 claims 1, 4-14, 18-20, 28-29, and 33-50.
This is a provisional nonstatutory double patenting rejection.
Response to Applicant’s Arguments
Applicant’s arguments filed on 11/21/2025 with respect to claims 1, 3, and 5-20 have been considered but are moot because the new matter and new ground of rejection does not rely on only prior applied reference but also include new references as prior arts of record for any teaching or matter specifically challenged in the argument.
Applicant’s argument:
The instant claims 1, 3 and 5-20 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 4-14, 18-20, 28, 29 and 33-50 of copending Application No. 18/247,161 in view of Gehrke et al. (U.S. Publication No. 2018/0372755 A1, hereinafter, "Gehrke"), and further in view of Liu et al. (Microb Cell Fact (2017) 16:223, hereinafter, "Liu"), Ning et al. (Virologica Sinica, 2017, 32 (1), p. 3-15, hereinafter, "Ning"), Zhang et al. (Sci Rep 6, 25856, p. 1-15, hereinafter, "Zhang"), Bornholdt et al. (Science. 2016 Mar 4;351(6277):1078-83, hereinafter, "Bornholdt"), Szunerits et al. (Nanomaterials (Basel). 2020 Jun 29; 10(7): 1271, hereinafter, "Szunerits"), Wu et al. (ACS Applied Nano Materials, 3(10), 9560-9580, hereinafter, "Wu"), Hung et al. (Nanomedicine. 2014 May;10(4):819-29, hereinafter, "Hung"), Alejo-Canchol et al. (PLoS Negl Trop Dis. 2020 Feb 18;14(2) , hereinafter, "Alejo-Canchol") and Koh et al. (Sensors 2009, 9, 8130-8145, hereinafter, "Koh").
For the same reasons presented above with respect to the rejections under 35 U.S.C. § 103, the cited art fails to teach or suggest each and every element of claim 1, 18, or 19, particularly as amended to include the amino acid sequence information of the CDRs of the recited sGP49 and sGP7 nanobodies. Therefore, claims 1, 3 and 5-20 are not obvious over claims 1, 4-14, 18-20, 28, 29 and 33-50 of copending Application No. 18/247,161 in view of the referenced cited art. Accordingly, the Applicant respectfully requests that this rejection be withdrawn.
In Response: Applicant’s arguments filed on 11/21/2025 with respect to claims 1, 3, and 5-20 as provisionally rejected on the ground of nonstatutory double patenting have been considered but are not persuasive in view of new ground of rejection the arguments are moot, and the rejections are maintained.
Response to Applicant’s Remarks filed on 02/20/2026
24. Applicant’s remarks/arguments filed on 02/20/2026:
Claims 1, 3 and 5-20 are currently pending in the application. Claims 1, 18, and 19 were amended in the response to the Non-Final Office Action filed November 21, 2025. As explained throughout the Rule 132 Declaration of Chao Wang accompanying that response to the Non- Final Office Action filed November 21, 2025, including, for example, in Section 9.4 thereof, "the inclusion of SEQ ID NOs:1-6 in the claims and sequence listing does not introduce new matter, but instead provides a formal record of nanobody sequences already described and experimentally supported in the application as filed."
These materials demonstrate that the inventors were in possession of the CDR sequences of the sGP49 and sGP7 nanobodies at the time of filing and that the application, as filed, enables one of ordinary skill in the art to make and use the claimed nanobodies without undue experimentation.
In connection with the Sequence Listing submitted concurrently herewith, the undersigned hereby states that the submission, in accordance with 37 C.F.R. §1.825(a)(4), does not include new matter. Applicant asserts that the listing is now in compliance with 37 CFR §§ 1.821-1.825.
25. Relevant Prior Art:
(i) Butterfield et al 2019 (WO 2019094669 A2, 05/16/2019 with an earlier priority to PCT/US18,59943, filed 11/09/2018) discloses SEQ ID NO: 591, an antibody CDR sequences that has 100% amino acid identity with instant claimed sGP7 nanobody CDR 1-3 SEQ ID NOs: 1-3.
Qy 1 FSSSNYA-------------------RWDNVKA--------------------------- 13
||||||| |||||||
Db 27 FSSSNYAMGWFRQAPGKEREFVSAISRWDNVKAYYADSVKGRFTISRDNSKNTVYLQMNS 86
Qy 14 -----------AMVDDYWDPG 23
||||||||||
Db 87 LRAEDTATYYCAMVDDYWDPG 107
(ii) Nguyen et al 2018 (US20180141998A1, 05/24/2018 priority to US15/568,487 filed on 04/25/2016) disclosed SEQ ID NO: 1208763 that has 100% identity with instant SEQ ID NO: 5 CDR2 of instant claimed sGP49 nanobody.
Qy 1 RSSSAAD 7
|||||||
Db 1 RSSSAAD 7
(iii) Hultberg et al 2015 (US20110182897A1, 07/28/2011, patent granted US9193780B2, 11/24/2025) teaches treatment of viral diseases including Ebola virus using nanobody. The present invention is in its broadest sense also not particularly limited to or defined by a specific envelope protein of a virus or a specific class, category or type of envelope proteins of a virus against which the amino acid sequences and polypeptides of the invention are directed. For example, the amino acid sequences and polypeptides may be directed against any envelope protein of a virus, e.g. the GP2 protein of Ebola virus (See, para [0058], [0209]-[0210], [0844]). The Nanobodies and polypeptides of the invention are generally directed against any epitope e.g. the GP2 protein of Ebola virus (See, para [0844]). Hultberg et al 2015 teaches SEQ ID NO: 2580 that has 92.7% identity match (with 1 conservative amino acid substitution) with instant claimed sGP49 CDR 1 SEQ ID NO: 4.
Query Match 92.5%; Score 37; Length 128;
Best Local Similarity 85.7%;
Matches 6; Conservative 1; Mismatches 0; Indels 0; Gaps 0;
Qy 1 DTYYIYA 7
||||||:
Db 101 DTYYIYS 107
(iv) Park et al 2021 (US11123402B1, 09/21/2021) discloses SEQ ID NO: 1 that has that has 52% identity match (with some conservative amino acid substitution) with instant claimed sGP49 CDR 3 SEQ ID NO: 6. Query Match 52.0%; Score 51; Length 559;
Best Local Similarity 50.0%;
Matches 7; Conservative 6; Mismatches 1; Indels 0; Gaps 0;
Qy 1 SWHVRFDDYEKVIS 14
:||| ||::|:::|
Db 285 AWHVGFDNFEQLLS 298
(v) Kang et al 2019. COMBINES-CID: An Efficient Method for De Novo Engineering of Highly Specific Chemically Induced Protein Dimerization Systems. J Am Chem Soc. 2019 Jul 17;141(28):10948-10952. Kang et al 2019 teaches chemically induced dimerization (CID) systems, in which two proteins dimerize only in the presence of a small molecule ligand, and developed a combinatorial binders-enabled selection of CID (COMBINES-CID) method broadly applicable to different ligands. Demonstrated a proof-of-principle by generating nanobody. We applied the CID system to a sensitive sandwich enzyme-linked immunosorbent assay-like assay of cannabidiol in body fluids with a detection limit of ∼0.25 ng/mL. COMBINES-CID provides an efficient, cost-effective solution for expanding the biosensor toolkit for small molecule detection.
Esmagambetov et al 2021. Nanobodies Are Potential Therapeutic Agents for the Ebola Virus Infection. Acta Naturae. 2021 Oct-Dec;13(4):53-63. (Year: 2021).
Bu et al 2026. A highly potent and broadly accessible bispecific nanobody for the treatment of ebola virus infections. PLoS Pathog. 2026 Jan 27;22(1):e1013878. (Year: 2026).
Conclusion
26. No claim is allowed.
27. Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
28. A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/SAMADHAN JAISING JADHAO/Examiner, Art Unit 1672
/BENNETT M CELSA/Primary Examiner, Art Unit 1600