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 .
Priority
The present application was filed on 11/11/2022. This application claims benefit of U.S. Provisional Patent application 63/295,101 filed 12/30/2021.
Election/Restrictions
Applicant did not elect a single species in species groups A, and B, and C as requested in the Requirement for Restriction/Election. However, upon further consideration, Examiner has withdrawn the species election from the Requirement for Restriction.
Applicant’s election without traverse of Group II (claims 7-20) in the reply filed on 02/27/2026 is acknowledged.
Claim Status
Claims 1-6 are withdrawn. Claims 7-17 are amended. Claims 7-20 are pending and examined.
Drawings
The drawings are objected to as failing to comply with 37 CFR 1.84(p)(5) because they include the following reference character(s) not mentioned in the description: “anti-His” in figures 1C, 3-5, 7-9, and 13-14.
Corrected drawing sheets in compliance with 37 CFR 1.121(d), or amendment to the specification to add the reference character(s) in the description in compliance with 37 CFR 1.121(b) are required in reply to the Office action to avoid abandonment of the application. Any amended replacement drawing sheet should include all of the figures appearing on the immediate prior version of the sheet, even if only one figure is being amended. Each drawing sheet submitted after the filing date of an application must be labeled in the top margin as either “Replacement Sheet” or “New Sheet” pursuant to 37 CFR 1.121(d). If the changes are not accepted by the examiner, the applicant will be notified and informed of any required corrective action in the next Office action. The objection to the drawings will not be held in abeyance.
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.
Claims 7-20 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.
Claim 7 recites “the patient” and “the mild/moderate…case” in lines 2-3. There is insufficient antecedent basis for this limitation in the claim because a “patient” and a “mild/moderate…case” have not been recited earlier in the claim. Also, it is unclear if “the patient” is one of subjects recited later in the claim, and it is unclear which subject (e.g., a first, or a second, or a third subject) in the claim is referred to “the patient”.
Claim 7 recites “SARS-CoV-2 variant” in page 3, “a virus” and a variant of the virus” in page 4. It is unclear if a virus or variant of virus in page 4 is SARS-CoV-2.
Claim 7 recites “SARS-CoV-2 variant”, claim 13 recites ““SARS-CoV-2 virus”. It is unclear which is SARS-CoV-2 virus and which is SARS-CoV-2 variant in the claim.
Claim 7 recites “in vitro detecting an immune response in a second subject after vaccination”. It is unclear what vaccine was used for the second subject, so it is unclear if the immune response in the second subject is for SARS-CoV-2 variant.
Claim 7 recites “an extracellular domain” on page 4 line 3 and “an extracellular region” on page 4 line 4. It is unclear how the domain and the region differ from each other. On page 4 line 4, claim 7 recites “an extracellular region … of a spike protein from a variant of the virus”, on page 4 line 10, claim 7 recites “the extracellular domain of the spike protein of the variant of the virus”. There is insufficient antecedent basis for “the extracellular domain of the spike protein of the variant of the virus” because it is an extracellular region, but not an extracellular domain, of the spike protein of the variant of the virus which has been recited earlier in the claim.
Claim 7 recites “A method of using a protein microarray for categorizing the patient into the mild/moderate, severe, or critical case, comprising: in vitro detecting a protective efficacy of a vaccine, an antibody drug, or a small molecule drug against SARS-CoV-2 variant infection in a first subject; in vitro detecting an immune response in a second subject after vaccination, or in vitro detecting an immune response in a third subject after being infected with the SARS-CoV-2 variant;…” However, the claim does not recite any method step to take the subject matter of the claim except the protein microarray. For example, it is unclear how to detect a protective efficacy of a vaccine, or an antibody drug, or a small molecule drug against SARS-CoV-2 variant infection: what parameter is used to define the protective efficacy of a vaccine, or an antibody drug, or a small molecule drug, e.g., a parameter may be a level of IgG or IgM antibody response to a COVID-19 vaccine; and how to use the protein microarray to detect a protective efficacy of a vaccine, or an antibody drug, or a small molecule drug. It is unclear how to define a protective level of the parameter, e.g., at which level of IgG or IgM is considered a protective level. Also, it is unclear what immune response needs to be detected. It is unclear how to do the immune response detections by using the claimed protein microarray. It is unclear how to categorize the patient after detecting a protective efficacy of a vaccine, or detecting an immune response in the subjects.
Claim 11 recites “ the same brand name or different brand names”. There is insufficient antecedent basis for the “brand name” because a word “brand name” has not been recited in previous claims.
Claim 17 recites “an extracellular domain” and “an extracellular region” on page 6 lines 17-18. It is unclear how the domain and the region differ from each other. On page 6 line 18, claim 17 recites “an extracellular region … of a spike protein from a variant of the virus”, on page 6 line 24, claim 17 recites “the extracellular domain of the spike protein of the variant of the virus”. There is insufficient antecedent basis for “the extracellular domain of the spike protein of the variant of the virus” because it is an extracellular region, but not an extracellular domain, of the spike protein of the variant of the virus which has been recited earlier in the claim.
Claim 17 recites “the plurality of protein array blocks.” There is insufficient antecedent basis for “protein array blocks” because “protein array blocks” have not been recited earlier in the claim.
Claim 17 recites “a first fluorescently labeled angiotensin-converting enzyme 2 (ACE2) receptor on a human cell surface.” It is unclear if the first labeled reagent is: 1/ a human cell expressing an ACE2 receptor on its surface and the whole cell is labeled; or 2/ a human ACE2 receptor is labeled and used as the first labeled reagent.
Claims 18 and 19 recite “the method according to any one of claim 17”. It is unclear what “any one of claim 17” means.
All dependent claims are also rejected based on their dependency of the defected parent claims.
Claim Rejections - 35 USC § 101
35 U.S.C. 101 reads as follows:
Whoever invents or discovers any new and useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof, may obtain a patent therefor, subject to the conditions and requirements of this title.
Claims 7-16 are rejected under 35 U.S.C. 101 because the claimed invention is directed to an abstract idea without significantly more. The claims recite the method of disease severity assessment which is judicial exception (e.g., abstract idea). This judicial exception is not integrated into a practical application because there is no practical application recited in the claims. The claims do not include additional elements that are sufficient to amount to significantly more than the judicial exception because the additional steps amount to mere data gathering that does not go beyond well-understood, routine, and conventional activity.
Step 1 – Whether a claim is to a statutory category - YES
Claims 7-16 are directed a method of using a protein microarray for categorizing the patient into the mild/moderate, severe, or critical case of categorizing the patient into the mild/moderate, severe, or critical case, comprising: in vitro detecting a protective efficacy of a vaccine, an antibody drug, or a small molecule drug against SARS-CoV-2 variant infection in a first subject; in vitro detecting an immune response in a second subject after vaccination, or in vitro detecting an immune response in a third subject after being infected with the SARS-CoV-2 variant. Therefore, the instantly claimed invention falls into one of the four statutory categories.
Step 2A Prong 1 – Whether the claim is directed to a judicial exception (i.e. Does the claim recite an abstract idea, law of nature, or natural phenomenon?) – YES
As explained in MPEP § 2106.04(II), a claim “recites” a judicial exception when the judicial exception is “set forth” or “described” in the claim.
Claim 7 recites a method of categorizing the patient into the mild/moderate, severe, or critical case by detecting an immune response in a patient who is infected with the SARS-CoV-2 or is vaccinated.
The broadest reasonable interpretation of claim 7 of “categorizing” is abstract idea, specifically, abstract mental processes. The “categorizing” step involves assessing the level of immune response in a sample from a patient, then making an evaluation or judgment as if the patient is mild/moderate, or severe, or critical. The step could be performed in the human mind, or by a human using pen and paper, insofar as it reads on comparing level of immune response among patients and drawing conclusions from this about the severity of the disease.
Moreover, the “categorizing” step can also be regarded as a law of nature, namely, the naturally occurring correlation between level of immune response and the severity of illness.
Thus, the claim 7 falls into judicial exception.
Step 2A Prong 2 - Does the claim recite additional elements that integrate the judicial exception into a practical application? NO
The Step 2A, Prong 2 analysis requires identifying whether there are any additional elements recited in the claim beyond the judicial exception(s), and evaluating those additional elements to determine whether they integrate the exception into a practical application of the exception.
Claims 7-16 do not recite any additional element that integrate the exception into a practical application of the exception. The additional steps of describing a structure of the protein microarray, administering a vaccine, and treating a patient are insufficient to integrate the exception into a practical application because the purpose is merely to obtain data.
There is no subsequent step recited after the “categorizing” step that would practically apply the method depending on the results of the detecting immune response from a patient, e.g., treatment or other process steps that are performed after determining the severity of the patient.
Step 2B: Whether the additional elements contribute an “inventive concept”
In the second step it is determined whether the claimed subject matter includes additional elements that amount to significantly more than the judicial exception. See MPEP 2106.05.
Briefly, the claims 7-16 do not include additional elements that are sufficient to amount to significantly more than the judicial exception because of the following reasons.
Simply appending well-understood, routine, conventional activities previously known to the industry, specified at a high level of generality, to the judicial exception, has been found to be insufficient to add “significantly more” (MPEP 2106.05(I)(A)).
In claim 7, the use of a protein microarray for detecting an immune response in a subject does not add a meaningful limitation to the instant method as it would have been routinely used by those of ordinary skill in the art. Moreover, claim 7 does not recite how to use the protein microarray for detecting an immune response in a subject, thus, the immune response detection could be performed by any technique known in the art including those disclosed in the instant specification.
The use of a protein microarray in detecting immune response is well-understood, routine, and conventional. This position is supported by Wohlstadter et al. (US20210349104) and Wang (US20220026428).
Wohlstadter teaches a method of using a protein microarray for detecting an immune response in a subject (see Abstract). The immunoassay method comprises: contacting the biological sample with a surface comprising a viral antigen in each binding domain on the surface, e.g., a protein microarray (see par.166). The viral antigen can be a receptor binding domain of spike protein from different SARS-CoV-2 strains (see par.166).
Wang teaches a method for detection of antibodies to a spike protein or fragment thereof of SARS-CoV (see Abstract, and par.17). The spike proteins or fragments thereof of SARS-CoV are immobilized on a solid support, i.e., a protein microarray (see Abstract, and par.17). The SARS-CoV may be SARS-CoV-2 or a variant thereof (see par.22). The spike protein or fragment may comprise or consist of the receptor binding domain (RBD) of the spike protein.
Therefore, the claims are not drawn to eligible subject matter as they are directed to an abstract idea and a law of nature without significantly more.
Claim Rejections - 35 USC § 103
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.
Claims 7-8, 10, 12-13 and 16 are rejected under 35 U.S.C. 103 as being unpatentable over Wohlstadter et al. (US20210349104) in view of Wang (US20220026428).
For claim 7, Wohlstadter teaches a method of using a protein microarray for detecting an immune response in a subject (see Abstract). The method comprises:
the protein microarray comprising: a substrate comprising a plurality of protein array blocks on a surface of the substrate (see par.166: teaching “the multiplexed immunoassay method comprises: contacting the biological sample with a surface comprising a viral antigen in each binding domain on the surface… the surface comprises a multi-well assay plate, wherein each well comprises ten distinct binding domains);
wherein the at least two proteins comprise an extracellular domain or a receptor binding domain of a spike protein from a virus, and an extracellular region or a receptor binding domain of a spike protein from a variant of the virus (see pars. 99, 150 and 166: teaching the viral antigen in each binding domain is S-RBDs (which is receptor binding domain of spike protein) from different SARS-CoV-2 strains, e.g., wild-type, B.1.1.7, 501Y.V2 etc.) This teaching encompasses that the at least two proteins in the protein array are the S-RBD of a virus and the S-RBD of a variant of the virus.
Wohlstadter teaches that the method is adopted for COVID-19 research, epidemiology, and vaccine development (see par.79). The method of detecting immunoglobulin(s) which conducted on non-bodily samples or bodily samples (e.g., serum, plasma, saliva) aids in assessing human immune responses to COVID-19 infection and vaccination (see par.79).
Wohlstadter does not teach the receptor binding domain of the spike protein of the virus comprises an amino acid sequence of SEQ ID NO: 10, or the receptor binding domain of the spike protein of the variant of the virus comprises any one of an amino acid sequence of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, or any combination thereof.
Wang teaches a method for detection of antibodies to a spike protein or fragment thereof of SARS-CoV (see Abstract, and par.17). The spike proteins or fragments thereof of SARS-CoV are immobilized on a solid support (see Abstract, and par.17). The SARS-CoV may be SARS-CoV-2 or a variant thereof (see par.22). The spike protein or fragment may comprise or consist of the receptor binding domain (RBD) of the spike protein. Wang teaches that the receptor binding domain of the spike protein of the virus comprises an amino acid sequence of SEQ ID NO: 10 (see par.23: teaching that the spike protein or fragment may comprise or consist of an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NOs: 13 or 26; which SEQ ID NO: 13 of Wang is 100% sequence identity to the nucleotide sequence of SEQ ID NO: 10 of the instant application (see result in the table below)).
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790
1865
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Greyscale
Sequence search result excerpted from ABSS
Wang teaches that the variant of SARS-CoV may comprise a nucleotide sequence having at least 80% sequence identity to the nucleotide sequence with SARS-CoV (see par.160). Therefore, the teaching of Wang encompasses the receptor binding domain of the spike protein of the variant of the virus comprising any one of an amino acid sequence of SEQ ID NO: 11 (which is 98.9% sequence identity to the nucleotide sequence with SARS-CoV SEQ ID NO: 13 of Wang), or SEQ ID NO: 12 (which is 98.2% sequence identity to the nucleotide sequence with SARS-CoV SEQ ID NO: 13 of Wang), or SEQ ID NO: 13 (which is 99.3% sequence identity to the nucleotide sequence with SARS-CoV SEQ ID NO: 13 of Wang), or SEQ ID NO: 14 (which is 98.2% sequence identity to the nucleotide sequence with SARS-CoV SEQ ID NO: 13 of Wang), or SEQ ID NO: 15 (which is 99.3% sequence identity to the nucleotide sequence with SARS-CoV SEQ ID NO: 13 of Wang), or SEQ ID NO: 16 (which is 98.6% sequence identity to the nucleotide sequence with SARS-CoV SEQ ID NO: 13 of Wang), or SEQ ID NO: 17 (which is 98.2% sequence identity to the nucleotide sequence with SARS-CoV SEQ ID NO: 13 of Wang).
Wang also teaches that the method is adopted for many aspects of COVID-19 investigation from contact tracing, seroprevalence survey, reservoir/intermediate animal tracking to assessment of herd immunity, longevity of protective immunity and efficacy of different vaccine candidates (see par.387).
Therefore, 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 Wohlstadter, by coating the receptor binding domains of the spike protein from SARS-CoV-2 and its variants as taught by Wang to assess human immune responses to COVID-19 infection and to assess the efficacy of covid-19 vaccination or covid-19 drug testing. By doing that, the method can apply for COVID-19 investigation from immune response to assessment of herd immunity, longevity of protective immunity and efficacy of different vaccine candidates as taught by Wohlstadter and Wang as discussed above.
One having an ordinary skill in the art would have had a reasonable expectation of success in combining the prior art references because Wohlstadter and Wang are directed to the method of detecting neutralizing antibodies to SARS-CoV and its variants, wherein Wohlstadter is generic to the receptor binding domain of spike protein from different SARS-CoV-2 strains; Wang teaches the specific amino acid sequences of the receptor binding domain of spike protein from SARS-CoV-2 and the variants thereof. The combined teaching of Wohlstadter and Wang encompasses all the limitations of claim 7, thereby encompassing the method of using the protein microarray for categorizing the patient into the mild/moderate, severe, or critical case.
For claim 8, Wohlstadter and Wang teach the method according to claim 7, wherein the vaccine is a COVID-19 vaccine (see Wohlstadter at least pars.79 and 520: teaching that the method to detect immunoglobulin(s) conducted on non-bodily samples or bodily samples (e.g., serum, plasma, saliva) aids in assessing human immune responses to COVID-19 infection and vaccination).
For claim 10, Wohlstadter and Wang teach the method according to claim 8, wherein the first subject receives one dose of the COVID-19 vaccine (see Wohlstadter par.273: teaching that the biological sample is obtained from a subject prior to be administered with a vaccine).
For claim 12, Wohlstadter and Wang teach the method according to claim 7, wherein the antibody drug is a monoclonal antibody drug. Wohlstadter teaches that the biological sample is from the subject who is in drug research and development to better understand disease progression and infection lethality (see Wohlstadter pars.134 and 250). The drug is a monoclonal antibody drug (see Wohlstadter at least pars.49-50, 134, 575, 701, and 705).
For claim 13, Wohlstadter and Wang teach the method according to claim 12, wherein the monoclonal antibody drug is a monoclonal antibody against a spike protein of a SARS-CoV-2 virus, a monoclonal antibody against a S l domain of the spike protein of the SARS-CoV-2 virus, or a monoclonal antibody against a nucleocapsid protein of the SARS-CoV-2 virus.
Wohlstadter teaches detecting a neutralizing monoclonal antibody against the S proteins (or RBD fragment) from SARS-CoV and SARS-CoV-2 by using a neutralization serology assay (see Wohlstadter at least pars.29, 49, 701 and 711).
For claim 16, Wohlstadter and Wang teach the method according to claim 7, wherein the immune response comprises a human immunoglobulin G (IgG), a human immunoglobulin A (IgA), a human immunoglobulin M (IgM), or any combination thereof generated in the second subject and the third subject. (See Wohlstadter par.202: teaching that the biomarker is an antibody or antigen-binding fragment thereof, e.g., IgG, IgA, IgM etc.)
Claims 9 and 11 are rejected under 35 U.S.C. 103 as being unpatentable over Wohlstadter et al. in view of Wang, as applied to claim 7, and further in view of Khehra et al. (Tozinameran (BNT162b2) Vaccine: The Journey from Preclinical Research to Clinical Trials and Authorization, AAPS PharmSciTech (2021) 22: 172).
For claim 9, Wohlstadter and Wang teach the method according to claim 8. Wohlstadter is generic about the Covid-19 vaccine for immunogenicity assessments (see par.79), e.g., vaccine in Operation Warp Speed project or all funded Phase III clinical trials of vaccines. Wohlstadter does not teach a specific vaccine, e.g., BNTI62b2 vaccine (Pfizer-BioNTech), mRNA-1273 vaccine (Moderna), AZD1222 vaccine (Oxford/AstraZeneca), or JNJ-78436735 vaccine (Johnson & Johnson).
Khehra teaches that vaccine BNT162b2 from Pfizer-BioNTech is one of vaccine candidate in Operation Warp Speed. Khehra also teaches that vaccination development and production was an essential question for the prevention and global control of COVID-19. See Abstract. Khehra teaches that the assessing immune response in a subject after vaccination is one of the criteria to test the efficacy of vaccine. See Phase 1/2 Clinical Trials: Antibody Response on pages 2-3.
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to use the modified method of Wohlstadter to assess an immune response in a subject after any Covid-19 vaccination (e.g., BNT162b2) to test the efficacy of vaccine for the benefit of preventing and controlling Covid-19 as taught by Khehra. Moreover, Wohlstadter teaches that immunogenicity assessments should be done on vaccines in Operation Warp Speed project, so it would be obvious to do immunogenicity assessment on BNT162b2 because BNT162b2 is a vaccine candidate in Operation Warp Speed as taught by Khehra.
For claim 11, Wohlstadter and Wang teach the method according to claim 10. Wohlstadter does not teach that the subject gets more than one dose of Covid-19 vaccine.
Khehra teaches that the vaccine trial can be done with a single dose or multiple doses from the same vaccine (see page 2 Preclinical Trials and Preclinical Trials: Safety and Efficacy, see pages 2-3 Phase 1/2 Clinical Trials: Antibody Response).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to use the modified method of Wohlstadter to assess an immune response in a subject after any Covid-19 vaccination (e.g., BNT162b2), either with single dose or two doses, to test the efficacy of vaccine for the benefit of preventing and controlling Covid-19 as taught by Khehra. Moreover, one would have been motivated to assess a protective efficacy of more than one dose Covid-19 vaccination because a dual-dose regimen may be required to increase antibody titers (see Khehra page 3 col.1) which implies the increased protective efficacy of a vaccine.
Claim 14 is rejected under 35 U.S.C. 103 as being unpatentable over Wohlstadter et al. in view of Wang, as applied to claim 7, and further in view of Kai (Interactions of coronaviruses with ACE2, angiotensin II, and RAS inhibitors—lessons from available evidence and insights into COVID-19, Hypertension Research volume 43, pages648–654 (2020).
For claim 14, Wohlstadter and Wang teach the method according to claim 7. Wohlstadter does not teach that the small molecule drug comprises a receptor blocker.
Kai teaches that angiotensin-converting enzyme 2 (ACE2) has been shown to be a functional receptor for SARS-CoV-2 to enter host target cells (see Abstract). In addition to vaccine development and antiviral agents, the modulation of the ACE2, e.g., using angiotensin receptor blockers (ARB), is highlighted as a potential therapeutic target for COVID-19. See Potential therapies targeting ACE2 and angiotensin II. However, besides the beneficial effects of ARB in blocking the virus from entering its host cells, the use of ARB in SARS-CoV-infected patients causes a concern because ARB can upregulate ACE2 expression as shown in animal studies, thereby increasing the morbidity and mortality of COVID-19 (see Abstract). While there is no clinical or experimental evidence supporting that ARBs and ACEIs either augment the susceptibility to SARS-CoV-2 or aggravate the severity and outcomes of COVID-19 at present, further research needs to be done (see Abstract).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to use the modified method of Wohlstadter to assess a protective efficacy of a receptor blocker, e.g., ACE2 ARBs, to solve the question of whether ARBs and ACEIs have neutral, favorable, or harmful effects on the susceptibility to SARS-CoV-2 and the severity and outcomes of COVID-19 (see Kai Risk limits of RAS inhibitor therapy in the COVID-19 pandemic, par.4).
Claim 15 is rejected under 35 U.S.C. 103 as being unpatentable over Wohlstadter et al. in view of Wang, as applied to claim 7, and further in view of Kai (Interactions of coronaviruses with ACE2, angiotensin II, and RAS inhibitors—lessons from available evidence and insights into COVID-19, Hypertension Research volume 43, pages648–654 (2020), and Amat-Santos et al. (Ramipril in High-Risk Patients With COVID-19, Journal of the American College of Cardiology, Volume 76, Issue 3, 21 July 2020, Pages 268-276).
For claim 15, Wohlstadter and Wang teach the method according to claim 7. Wohlstadter does not teach that the small molecule drug comprises a receptor blocker, e.g., Perindopril or Ramipril.
Kai teaches the motivation of assessing the risk and benefit of an ACE2 receptor blocker in treating SARS-CoV-2 infected patients. See discussion in claim 14 above.
Amat-Santos teaches that ramipril is one of ARBs. Amat-Santos analyzes whether RAAS inhibitors ramipril modify the risk for COVID-19 (see Abstract).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to use the modified method of Wohlstadter to do more research for a protective efficacy of a receptor blocker, e.g., ramipril as taught by Amat-Santos, to solve the question of whether ramipril has neutral, favorable, or harmful effects on the susceptibility to SARS-CoV-2 and the severity and outcomes of COVID-19 as suggested by Kai (see Kai Risk limits of RAS inhibitor therapy in the COVID-19 pandemic, par.4).
Claims 17-20 are rejected under 35 U.S.C. 103 as being unpatentable over Wohlstadter et al. (US20210349104) in view of Wang (US20220026428), as applied to claim 7, and further in view of Thermoscientific (Pierce™ Protein-Free Blocking Buffers, September 2021), and Feyzkhanova et al. (Development of a microarray‑based method for allergen‑specific IgE and IgG4 detection, Clin Proteom (2017) 14:1).
For claim 17, Wohlstadter and Wang teach the protein microarray according to claim 7.
The protein microarray comprises: a substrate comprising a plurality of protein array blocks on a surface of the substrate; and at least two proteins immobilized on each of the plurality of protein array blocks, wherein the at least two proteins comprise an extracellular domain or a receptor binding domain of a spike protein from a virus, and an extracellular region or a receptor binding domain of a spike protein from a variant of the virus, wherein the receptor binding domain of the spike protein of the virus comprises an amino acid sequence of SEQ ID NO: 10; and the receptor binding domain of the spike protein of the variant of the virus comprises any one of an amino acid sequence of SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, or any combination thereof. See the discussion of Wohlstadter and Wang in claim 7 above.
Wohlstadter teaches a method for detecting an immune response in a subject (see Abstract) comprising the steps of:
providing a protein microarray (see par.166: teaching “the multiplexed immunoassay method comprises: contacting the biological sample with a surface comprising a viral antigen in each binding domain on the surface… the surface comprises a multi-well assay plate, wherein each well comprises ten distinct binding domains, see pars.409-413);
adding a blocking reagent to the plurality of protein array blocks of the protein microarray for reaction to obtain a first protein microarray (see par.415: teaching that a blocking solution is added to the plate to reduce non-specific binding of the coating solution or the biotinylated binding reagent to the surface);
providing a to-be-tested sample from the subject, adding the to-be-tested sample to the first protein microarray for reaction (see pars.409-413: teaching that a sample comprising the antibody biomarker is added into the protein microarray surface in step (b)), and then washing to obtain a second protein microarray (see par.414);
Wohlstadter teaches that reacting for 50 to 70 minutes followed by washing to obtain a third protein microarray (see pars.414 and 417: in step (b), the plate is incubated for about 10 minutes to about 6 hours, or about 30 minutes to about 4 hours, or about 45 minutes to about 2 hours, or about 1 hour);
and reading an optical signal generated from the third protein microarray by a signal reader to quantify the anti-human immunoglobulin antibody (see par.413).
Wohlstadter provides a fluorescently labeled angiotensin-converting enzyme 2 (ACE2) receptor on a human cell surface or a fluorescently labeled anti-human immunoglobulin antibody; adds the fluorescently labeled ACE2 receptor on the human cell surface (see par.143 and Example 6B: the detection reagent comprises a labeled competitor of the biomarker ACE2) or the fluorescently labeled anti-human immunoglobulin antibody to the second protein microarray (see par.37 and Example 5B: teaching that labeled anti-IgG antibody was used to detect IgG in negative, early positive, and late positive SARS-CoV-2 patient samples).
Wohlstadter teaches the competitive serology assay which uses ACE2 as a competitor of the anti-SARS-CoV S protein antibodies for the antigen. ACE2 competes with the anti-SARS-CoV S protein antibodies in the COVID-19 patient samples, and binds to the SARS-CoV S proteins immobilized on the surface of the microarray (see par.707). Lower signal detected in the COVID-19 sera indicates inhibition of the interaction between the SARS-CoV-2 S protein and its cognate receptor, ACE2, in COVID-19 patient samples (see par.707). The competitive serology assay taught by Wohlstadter and the serology assay of the instant application in claim 17 are analogous in evaluating the neutralizing activity of the anti-SARS-CoV S protein antibodies in the COVID-19 patient samples by inhibition of interaction between the SARS-CoV-2 S protein and ACE2 receptor. Wohlstadter differs from the claimed method in detecting the signal of the labeled ACE2 receptor and labeled anti-human immunoglobulins in separate assay (see Example 12 and Fig.18). However, the results between different assay formats (which are an assay of detecting the binding of the SARS-CoV-2 S protein and the labeled ACE2 receptor and an assay of detecting the binding of the SARS-CoV-2 S protein and the labeled anti-human immunoglobulins) are conversely well-correlated.
Wohlstadter does not teach to add both fluorescently labeled ACE2 receptors on the human cell surface and fluorescently labeled anti-human immunoglobulins into the same assay to detect the signals from labeled ACE2 receptor and labeled anti-human immunoglobulins simultaneously. Wohlstadter does not teach that the blocking reagent is a non-protein blocking reagent.
Thermoscientific teaches a non-protein blocking reagent which can be used for blocking excess binding sites in immunoassays (e.g., the assay taught by Wohlstadter in pars.409-413). These blocking buffers reduce or eliminate many of the problems encountered with traditional protein-blocking reagents. Additionally, Protein Free blocking buffers are compatible with antibodies and avidin/biotin systems (e.g., the assay taught by Wohlstadter in pars.409-413). Protein-Free T20 blocking buffer contains the detergent Tween™-20, which improves blocking performance in many detection systems. See Product description in page 1.
Feyzkhanova teaches a multiplex immunoassay for detecting two analytes in the same assay (see Abstract). The two analytes were detected by Cy3 and Cy5 fluorescent labeled antibodies (see at least Abstract, page 3 col.2 par.2). Feyzkhanova supports the possibility of the simultaneous detection of two analytes using the corresponding Cy5 and Cy3 fluorescent dyes (see Mutual influence of Cy3‑ and Cy5‑labeled antibodies on the developing system on pages 5 and 7).
Therefore, 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 Wohlstadter, by using a protein free blocking buffer as taught by Thermoscientific for the benefit of improving blocking performance in the immunoassay taught by Wohlstadter. One having an ordinary skill in the art would have had a reasonable expectation of success in using the protein free blocking buffer of Thermoscientific in the assay of Wohlstadter because the buffer is compatible with antibodies and avidin/biotin systems (e.g., the assay taught by Wohlstadter in pars.409-413).
Therefore, 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 Wohlstadter, by adding multiple labeled detectors (both labeled ACE2 receptor and labeled anti-human immunoglobulins) into one assay, so multiple analytes can be detected simultaneously as taught by the method of Feyzkhanova. One having an ordinary skill in the art would have been motivated to use multiplex immunoassay to detect multiple analytes in one assay for the benefit of convenience, time-saving and validating the correlation of the analytes in the immune reaction instead of doing two single assays like Wohlstadter. One having an ordinary skill in the art would have had a reasonable expectation of success in measuring different signals from different labeled immunocomplexes because Feyzkhanova supports the measurement of multi-fluorescence signals in immunoassay.
For claim 18, Wohlstadter, Wang, Thermoscientific, and Feyzkhanova teach the method according to claim 17, wherein the immune response comprises a human immunoglobulin G (IgG), a human immunoglobulin A (IgA), a human immunoglobulin M (IgM), or any combination thereof generated in the second subject and the third subject. (See Wohlstadter par.202: teaching that the biomarker is an antibody or antigen-binding fragment thereof, e.g., IgG, IgA, IgM etc.)
For claim 19, Wohlstadter, Wang, Thermoscientific, and Feyzkhanova teach the method according to claim 17. Wohlstadter teaches that ACE2 receptors and anti-human immunoglobulins are labeled with fluorescence. Wohlstadter does not teach the fluorescence is Cy3 and Cy5.
Feyzkhanova teaches a multiplex immunoassay for detecting two analytes in the same assay (see Abstract). The two analytes were detected by Cy3 and Cy5 fluorescent labeled antibodies (see at least Abstract, page 3 col.2 par.2). Feyzkhanova supports the possibility of the simultaneous detection of two analytes using the corresponding Cy5 and Cy3 fluorescent dyes (see Mutual influence of Cy3‑ and Cy5‑labeled antibodies on the developing system on pages 5 and 7).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to substitute the fluorescent labels in the modified method of Wohlstadter for the Cy5 and Cy3 fluorescent dyes. One having an ordinary skill in the art would have been motivated to use the system that contained mixtures of antibodies labelled with different fluorescent dyes, e.g., Cy5 and Cy3, because the differences in the signals of the individually labelled antibodies and their mixture fit within the standard deviation of the analysis of the microarrays, which indicates the propriety of the selected developing system in multiplex immunoassay (see Feyzkhanova Mutual influence of Cy3‑ and Cy5‑labeled antibodies on the developing system on pages 5 and 7).
One having an ordinary skill in the art would have had a reasonable expectation of success in measuring different signals from different labeled immunocomplexes because Wohlstadter is generic in fluorescence labels used for detecting analytes and Feyzkhanova supports the possibility of the simultaneous detection of two analytes using the corresponding Cy5 and Cy3 fluorescent dyes.
For claim 20, Wohlstadter, Wang, Thermoscientific, and Feyzkhanova teach the method according to claim 17, wherein the to-be-tested sample comprises a serum or a plasma of the subject (see Wohlstadter par.79: teaching that serology assay embodiments were conducted on non-bodily samples or bodily samples (e.g., serum, plasma, saliva)).
Conclusion
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/CHAU N.B. TRAN/Examiner, Art Unit 1677
/BAO-THUY L NGUYEN/Supervisory Patent Examiner, Art Unit 1677 April 9, 2026