DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Restriction/Election
Applicant's election with traverse of claims 1-14 in the reply filed on 05/01/2026 is acknowledged. The traversal is on the ground(s) that the inventions share a special technical feature since the Examiner’s use of obvious analysis is not applicable in the lack of unity analysis. This is not found persuasive because (1) lack of unity can be determined “a posteriori” through a combination of prior art references that makes obvious the shared technical feature (see MPEP 1850 II), and (2) the Applicant’s argument regarding the obviousness analysis is not persuasive under the broadest reasonable interpretation of the claims which do not explicitly require the antigenic peptides to be separate molecules.
Claim 47 is withdrawn from further consideration pursuant to 37 CFR 1.142(b), as being drawn to a nonelected groups of inventions, there being no allowable generic or linking claim. Applicant timely traversed the restriction (election) requirement in the reply filed on 05/01/2026.
The requirement is still deemed proper and is therefore made FINAL.
However, while maintaining the restriction requirement for Group IV, upon reconsideration, the lack of unity for Group I, Group II, and Group III is withdrawn and claims 1-16, 27 and 29-31 are rejoined.
Claims Status
Amendments filed 07/26/2024 are entered.
Claims 47 are withdrawn pursuant to restriction requirement.
Claims 1-16, 27, 29, 30, and 31 are pending and under examination.
Priority
The application is a 371 application, filed 10/06/2023, of PCT application PCT/US2022/023811, filed 04/07/2022, which claims priority benefits from Provisional No. 63171638, filed 04/07/2021. The effective filing date of this application is 04/07/2021, the filing date of Provisional No. 63171638.
Information Disclosure Statement
The information disclosure statement(s) (IDS) submitted on 01/15/2025 is being considered by the examiner.
Nucleotide and/or Amino Acid Sequence Disclosures
REQUIREMENTS FOR PATENT APPLICATIONS CONTAINING NUCLEOTIDE AND/OR AMINO ACID SEQUENCE DISCLOSURES
Items 1) and 2) provide general guidance related to requirements for sequence disclosures.
37 CFR 1.821(c) requires that patent applications which contain disclosures of nucleotide and/or amino acid sequences that fall within the definitions of 37 CFR 1.821(a) must contain a "Sequence Listing," as a separate part of the disclosure, which presents the nucleotide and/or amino acid sequences and associated information using the symbols and format in accordance with the requirements of 37 CFR 1.821 - 1.825. This "Sequence Listing" part of the disclosure may be submitted:
In accordance with 37 CFR 1.821(c)(1) via the USPTO patent electronic filing system (see Section I.1 of the Legal Framework for Patent Electronic System (https://www.uspto.gov/PatentLegalFramework), hereinafter "Legal Framework") as an ASCII text file, together with an incorporation-by-reference of the material in the ASCII text file in a separate paragraph of the specification as required by 37 CFR 1.823(b)(1) identifying:
the name of the ASCII text file;
ii) the date of creation; and
iii) the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(1) on read-only optical disc(s) as permitted by 37 CFR 1.52(e)(1)(ii), labeled according to 37 CFR 1.52(e)(5), with an incorporation-by-reference of the material in the ASCII text file according to 37 CFR 1.52(e)(8) and 37 CFR 1.823(b)(1) in a separate paragraph of the specification identifying:
the name of the ASCII text file;
the date of creation; and
the size of the ASCII text file in bytes;
In accordance with 37 CFR 1.821(c)(2) via the USPTO patent electronic filing system as a PDF file (not recommended); or
In accordance with 37 CFR 1.821(c)(3) on physical sheets of paper (not recommended).
When a “Sequence Listing” has been submitted as a PDF file as in 1(c) above (37 CFR 1.821(c)(2)) or on physical sheets of paper as in 1(d) above (37 CFR 1.821(c)(3)), 37 CFR 1.821(e)(1) requires a computer readable form (CRF) of the “Sequence Listing” in accordance with the requirements of 37 CFR 1.824.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed via the USPTO patent electronic filing system as a PDF, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the PDF copy and the CRF copy (the ASCII text file copy) are identical.
If the "Sequence Listing" required by 37 CFR 1.821(c) is filed on paper or read-only optical disc, then 37 CFR 1.821(e)(1)(ii) or 1.821(e)(2)(ii) requires submission of a statement that the "Sequence Listing" content of the paper or read-only optical disc copy and the CRF are identical.
Specific deficiencies and the required response to this Office Action are as follows:
Specific deficiency - The Incorporation by Reference paragraph required by 37 CFR 1.821(c)(1) is missing or incomplete. See item 1) a) or 1) b) above.
The size of the ASCII text file must be in bytes, not kilobytes.
The date of creation on record is October 06, 2023 but the Incorporation by Reference of Sequence Listing statement recites October 05, 2023. The date of creation is the date of creation of the sequence listing, i.e. the electronic version of the sequence data that accompanies the application, and is the day the Applicant submits the sequence data to the Office and the Office receives it (automatically, when electronically submitted).
Required response – Applicant must provide:
A substitute specification in compliance with 37 CFR 1.52, 1.121(b)(3) and 1.125 inserting the required incorporation-by-reference paragraph, consisting of:
A copy of the previously-submitted specification, with deletions shown with strikethrough or brackets and insertions shown with underlining (marked-up version);
A copy of the amended specification without markings (clean version); and
A statement that the substitute specification contains no new matter.
Claim Rejections - 35 USC § 112
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.
Claim 3 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.
A broad range or limitation together with a narrow range or limitation that falls within the broad range or limitation (in the same claim) may be considered indefinite if the resulting claim does not clearly set forth the metes and bounds of the patent protection desired. See MPEP § 2173.05(c). In the present instance, claim 3 recites the broad recitation of at least 10 amino acids of the BP26 protein, and the claim also recites requiring progressively more of the BP26 protein, which is the narrower statement of the range/limitation. The claim(s) are considered indefinite because there is a question or doubt as to whether the feature introduced by such narrower language is (a) merely exemplary of the remainder of the claim, and therefore not required, or (b) a required feature of the claims.
Claim Rejections - 35 USC § 103
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 nonobviousness.
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.
Claim(s) 1, 10-16, 27, 29-31 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sun (Sun et al, CN112574320A, effective filing date 12/29/2020) in view of Wagner (Wagner et al, A fluorescent bead-based multiplex assay for the simultaneous detection of antibodies to B. burgdorferi outer surface proteins in canine serum; published 2011), Nolan (Nolan et al, Multiplexed and Microparticle-based Analyses: Quantitative Tools for the Large-Scale Analysis of Biological Systems; published 2006), and Wang (Wang et al, Evaluation of humoral and cellular immune responses to BP26 andOMP31 epitopes in the attenuated Brucella melitensis vaccinated sheep; published 2014).
Regarding claim 1, Sun teaches:
a kit (para. 61) for detecting immune responses to a Brucella canis infection (para. 6) in a mammal (para. 32) comprising:
a multiplex capture reagent comprising a first antigenic peptide from a Brucella canis outer membrane protein and a second antigenic peptide from a different outer membrane protein (para. 28 and 42);
a labeled detector against antibodies of the mammal (i.e. a HRP-labeled recombinant protein G) (para. 36) in an indirect ELISA (para. 32); and a solid substrate (i.e. ELISA plate) (para. 52).
In summary, Sun teaches an indirect ELISA (iELISA) method and kit for this method to detect Brucella canis infection in dogs. Sun teaches a fusion polypeptide comprising of antigenic peptides from outer membrane proteins BP26, Omp2b, Omp16, Omp25 and Omp31 from Brucella (para 28, 40, 42) is coated onto an ELISA plate and incubated with the test subject's (i.e. dog) serum (para. 35). The serum antibody that bound the antigenic polypeptide are then bound with an HRP-labeled protein G, which binds and labels the serum antibodies (para. 36).
Sun teaches the antigenic proteins are from Brucella and teaches this assay is to detect Brucella canis infections; however, Sun does not explicitly teach that the antigenic peptides are specifically from Brucella canis.
Sun does not explicitly teach the first antigenic peptide is linked to a first detectable label and the second antigenic peptide is linked to a second detectable label, i.e. that the two claimed antigenic peptides are (a) linked to a detectable label and (b) linked to different detectable labels from each other.
Sun teaches an HRP-labeled protein G that binds and labels antibodies, but does not explicitly teach a detection antibody.
However, Wagner teaches labeling of antigenic peptides (i.e. outer surface proteins of a bacteria) in a fluorescent bead-based multiplex assay for diagnosis of another pathogenic infectious bacteria in canine serum (Abstract).
Wagner teaches more than two antigenic peptides (Abstract) but they are individually linked to a detectable label (i.e. fluorescent beads) (p. 194, section: 2.4 Coupling of recombinant B. burgdorferi antigens to fluorescent beads), which serves as the solid support for the antigen. Wang teaches each antigenic peptide is linked to a fluorescent bead (i.e. each bead only has one type of antigenic peptide) but all the beads have the same fluorescence.
Wagner further teaches the use of a biotinylated rabbit anti-dog IgG(H+L) as a detection antibody (p. 192, section: 2.5. Luminex assay).
Sun and Wagner do not explicitly teach that the antigenic peptides are specifically from Brucella canis.
Sun and Wagner do not explicitly teach that the two antigenic peptides are linked to detectable labels that are different labels from each other.
However, Nolan teaches the advantages of a multiplex assay with different fluorescence-encoded microspheres (i.e. beads) (p. 319, col. 1, para. 3) including the ability to distinguish between proteins and quantify protein abundance (p. 319, col. 2, para. 3). Nolan further teaches that these beads can be used in immunoassays in which the antigen is displayed on beads and are used to detect and characterize antibodies circulating in blood, typically for detecting infectious disease (p. 320, col. 2, para. 3).
Further, Nolan teaches that many scientists will prefer to purchase assay kits for multiplexed assays that have been optimized and validated due to the time needed to set one up from scratch, motivating the development of a kit for this assay.
Sun, Wagner, and Nolan do not explicitly teach that the antigenic peptides are specifically from Brucella canis.
However, Wang teaches BP26 and OMP31 are the major periplasmic or outer membrane proteins of Brucella and both proteins are considered the diagnostic tools for antibody detection (p. 825, col. 1, para. 2), implying they are specific to Brucella, instead of generally present in several bacteria. Wang further teaches that the antigenic proteins BP26 and Omp31 are highly conserved within the Brucella genus, specifically including Brucella canis (Wang, page 825, Introduction). Wang further teaches that, in their sheep models infected with native membrane protein extracts, antibody levels gradually increased to a plateau at 2 months and stayed high for up to 7.5 months (p. 832, para. 1). Wang further teaches there are several common, species-specific, and individual animal specific epitopes on BP26 and Omp31 (Figure 3B). Wang further teaches that there are some overlapping epitope sequences and some epitope sequences unique to each protein (p. 832, para. 2).
It would have been obvious to one skilled in the art, before the effective filing date of the instant application, from the teachings of Wang that BP26 and OMP31 is conserved in Brucella canis also and therefore, the BP26 and OMP31 antigen for diagnosing Brucella canis in Sun is from Brucella canis.
Secondly, it would have been obvious to one skilled in the art, before the effective filing date of the instant application, the benefits of multiplexing with specifically BP26 and OMP31 in light of their diagnostic value, as taught by Wang, and the benefits of different detectable labels to be able to quantify protein levels, as taught by Nolan et al, especially since Wang teaches that the levels of antibodies to these antigens change over time throughout the infection. Further, since there are several epitopes on these antigens, as taught by Wang, the level of serum antibody for each antigen might vary. For example, OMP31 was not as good a diagnostic antigen as BP26 (Wang, p. 825, col. 2, para. 2) so it would be obvious to take into account the overall antibody response to both antigen in determining infection. Although the teachings of Wang are from sheep models, the variability in antigen response is reasonably likely applicable to other Brucella-infected subjects. Thirdly, it would have been obvious to one skilled in the art, before the effective filing date of the instant application, that the labeled Protein G, taught by Sun, and labeled rabbit anti-dog IgG, taught by Wagner, are alternatives to labeled antibody detectors. However, the rabbit anti-dog IgG(H+L), or antibodies against IgG of a specific species, is more specific to the desired target, which is desirable to mitigate false positives, in contrast to protein G binds IgG from many species.
One skilled in the art, before the effective filing date of the instant application, would be motivated to develop a kit to detect the immune response to Brucella canis infection using these two antigenic peptides specific to Brucella canis, and to further be able to quantify the level of each antigen to determine the level of immune response to each antigen as an indicator of infection and possibly even help characterize the stage of infection. One skilled in the art, before the effective filing date of the instant application, would be motivated to choose the antibodies against IgG of a specific species as the antisera detection moiety to mitigate false positive binding.
One skilled in the art, before the effective filing date of the instant application, would have reasonable expectation of success that this assay would diagnose Brucella canis infection because BP26 and OMP31 are known conserved antigens on Brucella, so the antisera of infected subjects would reasonably likely contain antibodies that bind to these antigens. Secondly, one skilled in the art would have reasonable expectation of success that the linkage of these antigens to detection labels, enabling the distinction of these two antigens, would integrate with the detection antibody label to enable quantification of each particular antigen, as Nolan teaches, for example, fluorescent beads and the detection antibody can be read by a conventional plate reader, like an ELISA. One would reasonably expect that each of the three different labels can be chosen to be distinguishable by their different emission wavelengths.
Claims 10-16, 27, 29-31 depend on claim 1. The teachings of the references regarding claim 1 are incorporated in its entirety for the dependent claims and discussed further below, as is relevant for each claim.
Regarding claim 10, as described above, Nolan teaches the use of two different fluorescent beads for each protein of interest. The reasons for incorporating this into the immunoassay for diagnosing Brucella canis infection is also described above.
Regarding claim 11, Wagner teaches the detection antibody is a biotinylated rabbit anti-dog IgG(H+L), which is an antibody with a detectable label (biotin), that is linked to the streptavidin-phycoerythrin, which provide a fluorescent signal (p. 192, section: Luminex assay). Under broadest reasonable interpretation, the detection antibody is linked to a detectable label that provides a fluorescent signal because the claims do not limit when the linkage occurs and does not limit how the fluorescent signal is linked to the detection antibody.
Regarding claim 12, Sun teaches Brucella canis infects dogs, but can also infect humans (para. 6); therefore, to detect the infection, the subject's serum is used, which would be dog serum (para. 35), and it would be obvious and motivated that human serum would be used if the subject was human.
Regarding claim 13, as previously described, Sun teaches the antigens are coated onto the surface of a multi-well plate (para. 33); therefore it uses a multi-well plate as the solid substrate.
Regarding claim 14, Sun teaches other Brucella canis proteins that could be used as an antigenic peptide, such as Brucella Omp2b, Omp16, and Omp25, in addition to the instantly claimed BP26 and Omp31.
It would have been obvious to one skilled in the art, before the effective filing date of the instant application, that more than two antigenic peptides can be included to detect Brucella canis infection, since there are more than two known diagnostic antigenic proteins on Brucella canis.
One skilled in the art, before the effective filing date of the instant application, would be motivated to create a multiplex assay to detect more than one antigenic peptides from Brucella canis since the antibody response could be to any of the antigenic proteins, and at varying levels. Therefore, detecting multiple epitopes can help to reduce false negatives.
One skilled in the art, before the effective filing date of the instant application, would have reasonable expectation of success in adding a third antigenic peptide for the same reasons of expected success for the two antigenic peptide system. Further, one would expect that adding a third antigenic peptides would be compatible with the system because there exists enough distinguishable detectable labels, such as fluorophores, to distinguish between the three antigenic peptides.
Claims 15-16 are directed to a method of using the kit of claim 1 for its purpose of detecting antigenic peptide from PB26 and Omp31 in subject's sample to detect Brucella canis infection. The teachings of how the combined references teach this method is previously provided in the discussion regarding claim 1. In summary, Sun teaches the foundational multiplex assay method to detect serum antibodies to multiple Brucella canis antigenic peptides to detect Brucella canis infection. Wagner teaches the antigenic peptides on separate solid substrates, specifically fluorescent beads, instead of as a fusion protein, as in Sun. Wang teaches BP26 and Omp31 in particular are diagnostic antigens for Brucella canis and motivates their selection for the assay. Nolan teaches multiplexed and microparticle-based quantitative analysis tools (Abstract). Nolan teaches the use of beads labeled with different fluorophores as the solid substrate for molecular analysis using flow cytometry (p. 319, section: Beads, dyes, instruments, and assays) and their use in multiplexed analysis of protein abundance. Nolan first teaches bead-based immunoassays generally employ the sandwich assay design in which different capture antibodies on distinct microspheres and reporter/secondary antibodies are used to simultaneously measure the levels of multiple analytes (p. 320, col. 2, para. 2).
As is further relevant to claims 15-16, Nolan then teaches a second class of immunoassays in which the antigen is displayed on the distinct beads and used to capture and characterize antibodies circulating in blood, which, in turn, are then detected with a labeled secondary antibody. This assay is used to detect exposures to infectious disease and monitor antigen-specific antibody responses to infection or vaccination (p. 320, col. 2, para. 3).
As is further relevant to claim 16, it is obvious that the use of different fluorophores on the beads and the labeled detection antibody is to enable the quantification of the amount of each antigen-specific antibody by measuring the beads that have a co-localized signal from the detection antibody and its own bead label.
Claims 27, 29, and 30 depend on claim 15. The teachings of the references regarding the parent claim are incorporated in its entirety for the dependent claims and discussed further below, as is relevant for each claim.
Regarding claim 27, as previously discussed, Nolan teaches the use of a flow cytometer to read the signals from the labeled beads/microparticle. Sun teaches that when the solid substrate is a plate, the signal can be read with a plate reader.
Regarding claim 29, as previously discussed, Nolan, for example, teaches the biological sample is blood or plasma (p. 320, col. 2, para. 3) to measure the antisera.
Regarding claim 30, as previously discussed, Sun and Wang teaches BP26 and Omp31 are two antigenic peptides that are used to diagnose Brucella canis infection, and therefore, positive detection of these antibodies specific to these antigens would be indicative of a Brucella canis infection.
Regarding claim 31, as previously discussed, the combined references provide a method of detecting and measuring the antigen-specific antibody at one timepoint. Wang teaches that the levels of antibodies to these antigens change over time throughout the infection, following a particular profile. For example, in their sheep models infected with native membrane protein extracts, antibody levels gradually increased to a plateau at 2 months and stayed high for up to 7.5 months (p. 832, para. 1).
It would have been obvious to one skilled in the art, before the effective filing date of the instant application, that the method to measure antigen-specific antibody at one timepoint can be repeated at different timepoints to arrive at the method of monitoring progression. Further, although the teachings of Wang are from sheep models, the time-dependent profile in antigen response is reasonably likely applicable to other Brucella-infected subjects.
One skilled in the art, before the effective filing date of the instant application, would be motivated to monitor the progression of the antigen-specific antibody as a measure of infection progression to monitor health risks and response to treatment.
One skilled in the art, before the effective filing date of the instant application, would have reasonable expectation of success that the method could be used at multiple timepoints by assaying biological samples taken from the different timepoints.
Claim(s) 11 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sun (CN112574320A, effective filing date 12/29/2020) in view of Wagner (Wagner et al, published 2011), Nolan (published 2006), and Wang (published 2014), as applied to claim 1, and further in view of Embers (Embers et al, Five-Antigen Fluorescent Bead-Based Assay for Diagnosis of Lyme Disease; published 2016).
Claims 11 depend on claim 1. The teachings of the references regarding claim 1 are incorporated in its entirety for the dependent claims and discussed further below, as is relevant for each claim.
Regarding claim 11 again, Embers teaches another multiplex immunoassay using antigenic peptides conjugated to fluorescent beads to detect a bacterial infection in mammal serum (Abstract and p. 295, col. 1, Section: Production of antigen-coupled cytometric beads), wherein the secondary (detection) antibody is conjugated to the fluorophore phycoerythrin, which provides a fluorescent signal (p. 295, col. 2, Section: Detection of specific antibodies and controls, para. 3). The samples were read on the Bio-Plex 200 suspension array system from Bio-Rad (p. 295, col. 2, Section: Detection of specific antibodies and controls, para. 3), which is designed to read the fluorescent beads in suspension through a high-throughput fluidics system, like a flow cytometer.
It would have been obvious to one skilled in the art, before the effective filing date of the instant application, that the fluorescent source could be directly linked to the secondary antibody, as in Embers instead of being directed to the secondary antibody, as in Wagner.
One skilled in the art, before the effective filing date of the instant application, would be motivated to minimize the number of required interactions to provide the fluorescent signal and therefore link the fluorophore directly to the secondary antibody.
One skilled in the art, before the effective filing date of the instant application, would have reasonable expectation of success since Ember teaches this as an alternative form of a secondary antibody that can provide a detectable fluorescent signal.
Claim(s) 2-4 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sun (CN112574320A, effective filing date 12/29/2020) in view of Wagner (Wagner et al, published 2011), Nolan (published 2006), Wang (published 2014) and Embers (published 2016), as applied to claim 1 and related claim 11 above, and further in view of Kim (Kim et al, Brucella Immunogenic BP26 Forms a Channel-like Structure; published 2013) and WP_002964581.1 (NCBI, outer membrane protein BP26/OMP28 [Brucella]; last updated 01/06/2020).
Claims 2-4 depend on claim 1, and claim 11 is a relevant embodiment of claim 1. The teachings of the references regarding claims 1 and 11 are incorporated in its entirety for the claims below and discussed further, as is relevant for each claim.
Regarding claim 2, Kim teaches the full 250 amino acid sequence for the BP26 protein, reported in the NCBI record WP_002964581.1, which has a 100% identity match to SEQ ID NO: 1.
Although Kim originally teaches this sequence from Brucella abortus, WP_002964581.1 was updated to teach that this sequence matches to multiple species of Brucella, including Brucella canis (see Image 1). To clarify terminology, Kim teaches that the transmembrane sequence of the OMP28 protein is cleaved away and the resulting protein is called BP26 (p. 1119, col. 2, para. 1), which is specifically sequence 29-250 of the full OMP28 protein, meaning that the transmembrane sequence is the N-terminal residues 1-28 (p. 1120, section: Results, para. 1 ). Therefore, what the instant calls the BP26 protein comprising of the amino acid sequence of BP26 is, more precisely the full-length OMP28 protein taught in Kim.
The sequence disclosed in WP_002964581.1 (denoted Db in the sequence alignment image, Image 2) has a 100% sequence identity to the entire query of SEQ ID NO: 1 (denoted Qy in Image 2).
Image 1:
PNG
media_image1.png
390
1213
media_image1.png
Greyscale
Image 2:
PNG
media_image2.png
480
630
media_image2.png
Greyscale
Regarding claim 3, Wang et al teaches several peptides along the BP26 protein, specifically, 11-16 amino acid peptides from the BP26 protein (Table 1 and p. 826, section: 2.3 Peptides and pools) and, of those, the antigenic (i.e. reactive) peptides are specifically listed in Table 2.
Regarding claim 4, Wang teaches antigenic peptides comprising a fragment of the BP26 protein without the N-terminal transmembrane domain of the BP26 protein, which are the BP26 variants starting from BP26-05 and onward (excluding residues 1-29, as seen in Table 1) that also appear in Table 2 (reactive peptides).
It would have been obvious and motivated to one skilled in the art, before the effective filing date of the instant application to use the whole or antigenic regions of the BP26 protein, outside the N-terminal transmembrane domain, as the capture reagent to identify BP26-specific antibody in test samples to detect infection by BP26-carrying Brucella canis.
One skilled in the art, before the effective filing date of the instant application, would have reasonable expectation of success since the antigenic regions encompassed in these claims are well characterized by Wang.
Claim(s) 5-7 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sun (CN112574320A, effective filing date 12/29/2020) in view of Wagner (Wagner et al, published 2011), Nolan (published 2006), Wang (published 2014), and Embers (published 2016), as applied to claims 1 and 11, and further in view of Vizcaino (Vizcaino et al, Cloning, Nucleotide Sequence, and Expression of the Brucella melitensis omp31 Gene Coding for an Immunogenic Major Outer Membrane Protein; published 1996) and Q93E87 (Q93E87, UniProt, Outer membrane protein Omp31 protein sequence; last updated 2019).
Claims 5-9 depend on claim 1, and claim 11 is a relevant embodiment of claim 1. The teachings of the references regarding claims 1 and 11 are incorporated in its entirety for the claims below and discussed further, as is relevant for each claim.
Regarding claim 5, Wang teaches the amino acid sequence for Brucella melitensis Omp31 (Table 1), which has a 100% sequence identity to the amino acid of SEQ ID NO: 2. Wang further teaches that Omp31 is expressed in B. canis (p. 825, col. 2, para. 2). Vizcaino also teaches the same Omp31 protein sequence as Wang (Figure 1) and this publication is linked to the sequence disclosure identified by UniProt identifier Q93E87.
Wang and Vizcaino do not explicitly teach the Brucella canis Omp31 protein sequence.
However, Q93E87 teaches the Omp31 sequence disclosed by Wang is also the predicted sequence for Brucella canis.
Regarding claim 6, Wang teaches 10-26 amino acid long reactive/antigenic peptide sequences of the Omp31 protein (p. 826, section: 2.3 Peptides and pools; Table 1 and Table 2).
Regarding claim 7, Wang and Q93E87 does not explicitly teach which region is the extracellular region of the Omp31.
However, Vizcaino, which teaches the same Omp31 protein sequence as Wang (Figure 1) and therefore Q93E87, teaches that the full-length Omp31 protein is a 240 amino acid protein and is cleaved between amino acid residue 19 and 20 to make the mature Omp31, which localizes to the bacteria surface (i.e. outer membrane), indicating that this region after residue 19 is the extracellular region (p. 3747, col. 1, para. 1 and col. 2, para. 2). In fact, Wang teaches peptides containing residues 1-21 do not induce a measurable antibody response but those that contain antigenic peptides after do induce an antibody response (i.e. are antigenic) (Table 1 and 2).
Regarding claim 5, it would have been obvious to one skilled in the art, before the effective filing date of the instant application that the whole Omp31 protein sequence of instant SEQ ID NO: 2, as disclosed by Wang and Q93E87, which contains multiple epitope sites, could be used to detect Omp31-specific antibodies.
Regarding claim 6, it would have been obvious to one skilled in the art, before the effective filing date of the instant application that a peptide of at least 10 amino acids of the Omp31 protein, comprising of specific epitopes, could also be used, since Wang teaches reactive peptides of at least 10 amino acids to determine the individual epitope regions.
Regarding claim 7, it would have been obvious to one skilled in the art, before the effective filing date of the instant application to use the extracellular region of the Omp31 protein and to know it was after residue 19 of the full length protein, because this is the region of the Omp31 that would be exposed to the immune system. Therefore, this is the region of the protein that the antibody response will recognize.
One skilled in the art, before the effective filing date of the instant application, would be motivated to use the whole or antigenic parts, which are in the extracellular region, of the Omp31 protein as the capture reagent to identify Omp31-specific antibody in test samples.
One skilled in the art, before the effective filing date of the instant application, would have reasonable expectation of success since the antigenic regions encompassed in these claims are well characterized by Wang.
Claim(s) 8 and 9 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sun (CN112574320A, effective filing date 12/29/2020) in view of Wagner (Wagner et al, published 2011), Nolan (published 2006), Wang (published 2014), Embers (published 2016), Vizcaino (Vizcaino et al, Cloning, Nucleotide Sequence, and Expression of the Brucella melitensis omp31 Gene Coding for an Immunogenic Major Outer Membrane Protein; published 1996), Q93E87 (Q93E87, Outer membrane protein Omp31 protein sequence; last updated 2019), as applied to claim 7 above, and further in view of Goldbaum (US 8685670 B2, published 2014).
Claims 8-9 depend on claim 7. The teachings of the references regarding claim 7 are incorporated in its entirety for the dependent claims and discussed further below, as is relevant for each claim.
Regarding claim 8, Goldbaum teaches an antigenic peptide of Omp31 disclosed in SEQ ID NO: 27 that comprises of part of instant SEQ ID NO: 8, yielding less than a 95% sequence match. It is missing the last three amino acids of SEQ ID NO: 8, as seen in the alignment below (Image 3: Qy = instant SEQ ID NO: 8 residues 1-27; Db = Goldbaum SEQ ID NO: 27).
Image 3:
PNG
media_image3.png
183
801
media_image3.png
Greyscale
However, Wang teaches the same antigenic peptide sequence taught by Goldbaum spanning across Wang's antigenic peptides OMP31-06 to OMP31-09 (screenshot of Wang, Table 1 below). Wang teaches that OMP31-09 is a reactive peptide but OMP31-10 is not, and that there are is an antibody response to OMP31-09 for a longer timescale than OMP31-08 (screenshot of Wang, Table 2 below).
Screenshot of Wang, Table 1:
PNG
media_image4.png
122
577
media_image4.png
Greyscale
Screenshot of Wang, Table 2:
PNG
media_image5.png
82
578
media_image5.png
Greyscale
It would have been obvious to one skilled in the art, before the effective filing date of the instant application, that the sequence of OMP31-10 is not an antigenic peptide sequence of OMP31, and therefore, it could be excluded from OMP31-09, leaving the highlight sequence, including the LDV residue. It would have been obvious that this highlighted region of OMP31-09 comprises of an antigenic region of OMP31 and therefore can be included into the antigenic peptide sequence taught by Goldbaum to arrive at instant SEQ ID NO: 8. The addition of this ending LDV sequence to the antigenic peptide taught by Goldbaum is further supported by the antibody response to OMP31-09 being maintained for a longer timescale than OMP31-08, which could be attributed to the additional DV sequence that OMP31-08 does not have. This stronger antigenicity to OMP31-09 motivates the detection of this sequence.
One skilled in the art, before the effective filing date of the instant application, would be motivated to include sequences that are likely highly antigenic, since these are what the antisera will react to.
One skilled in the art, before the effective filing date of the instant application, would have reasonable expectation of success that this sequence would be an antigenic sequence useful for the detection of Omp31-specific antibodies in Brucella canis infected subjects based on Goldbaum's disclosure of the antigenic peptide sequence that differs from instant SEQ ID NO: 8 by the last LDV sequence and the likely antigenic contribution of the additional LDV sequence that can be reasoned by the teachings of Wang.
Regarding claim 9, Goldbaum (US 8685670 B2, published 2014) teaches the antigenic peptide of Omp31 disclosed in SEQ ID NO: 27 (Db) that comprises of instant SEQ ID NO: 9 (Qy) in the sequence alignment below (Image 4).
Image 4:
PNG
media_image6.png
202
812
media_image6.png
Greyscale
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to BONIRATH CHHAY whose telephone number is (571)272-0682. The examiner can normally be reached Mon-Thu 8AM-5PM EST.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Bao-Thuy Nguyen can be reached at (571) 272-0824. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/BONIRATH CHHAY/Examiner, Art Unit 1645
Thursday, July 2, 2026
/BAO-THUY L NGUYEN/Supervisory Patent Examiner, Art Unit 1677 July 6, 2026
Prosecution Insights