DETAILED ACTION
Election/Restrictions
Applicant’s election of the species of nucleocapsid protein is acknowledged. Claims 1-3, 5-9, 11-19, 21-25, 27-31, 33-41, 43 and 44 are under examination. Claims 4, 10, 20, 26, 32 and 42 are withdrawn from consideration being directed to non-elected species.
Claims Summary
Claims 1-3, 5 and 6
Claim 1 is directed to a kit comprising:
A composition comprising:
A singlet oxygen-activatable chemiluminescent compound bound to a first target antigen, directly or indirectly; and
A fluorescent molecule excited by the compound;
A composition comprising:
A singlet oxygen-activatable chemiluminescent compound bound to a second target antigen, directly or indirectly; and
A fluorescent molecule that is excited by the compound, wherein the fluorescent molecule is different from the fluorescent molecule of (a) and emits light at a different wavelength than the fluorescent molecule of (a);
A biotinylated first target antigen, wherein the antigen is at least a portion of a SARS-CoV-2 spike protein; the antigen comprises an RBD of S1 protein (claim 2);
A biotinylated second target antigen, wherein the antigen is at least a portion of a SARS-CoV-2 protein other than the spike protein; the antigen comprises at least a portion of SARS-CoV-2 nucleocapsid (N) protein (claim 3); and
A composition comprising a sensitizer that generates singlet oxygen in its excited state and is bound to a biotin-specific binding partner, directly or indirectly; the sensitizer is a photosensitizer (claim 5); the biotin-specific binding partner is avidin, an analog of avidin, and an antibody against biotin (claim 5).
Claims 23-25, 27 and 28
Claim 23 is directed to a kit comprising:
A composition comprising:
A singlet oxygen-activatable chemiluminescent compound bound to a first target antigen, directly or indirectly; the antigen comprises an RBD of S1 protein (claim 24); and
A fluorescent molecule excited by the compound;
A composition comprising:
A singlet oxygen-activatable chemiluminescent compound bound to a second target antigen, directly or indirectly; wherein the antigen is at least a portion of a SARS-CoV-2 N protein (claim 25); and
A fluorescent molecule that is excited by the compound, wherein the fluorescent molecule is different from the fluorescent molecule of (a) and emits light at a different wavelength than the fluorescent molecule of (a);
At least one biotinylated anti-human Ig antibody; and
A composition comprising a sensitizer that generates singlet oxygen in its excited state and is bound to a biotin-specific binding partner, directly or indirectly; the sensitizer is a photosensitizer (claim 27); the biotin-specific binding partner is avidin, an analog of avidin, and an antibody against biotin (claim 27).
The intended use of the kits of claims 1-3, 5, 6, 23-25, 27 and 28 is for performing a multiplex assay that utilizes a chemiluminescent detection system for determining the presence and/or concentrations of multiple anti-SARS-CoV-2 antibodies in a sample. The singlet oxygen-activatable chemiluminescent compound of each of (a) and (b) is a substance that undergoes a chemical reaction with singlet oxygen to form a metastable intermediate species that can decompose with the simultaneous or subsequent emission of light (claims 5 and 27). The fluorescent molecules of (a) and (b) are each independently selected from terbium, uranium, samarium, europium, gadolinium and dysprosium (claims 6 and 28).
Claims 6-9 and 11-14
Claim 7 is directed to a microfluidics device comprising an inlet channel through which the samples is applied, and at least a first compartment capable of being in fluid communication with the inlet channel and containing:
A composition comprising:
A singlet oxygen-activatable chemiluminescent compound bound to a first target antigen, directly or indirectly; and
A fluorescent molecule excited by the compound;
A composition comprising:
A singlet oxygen-activatable chemiluminescent compound bound to a second target antigen, directly or indirectly; and
A fluorescent molecule that is excited by the compound, wherein the fluorescent molecule is different from the fluorescent molecule of (a) and emits light at a different wavelength than the fluorescent molecule of (a);
A biotinylated first target antigen, wherein the antigen is at least a portion of a SARS-CoV-2 spike protein; the antigen comprises an RBD of S1 protein (claim 8);
A biotinylated second target antigen, wherein the antigen is at least a portion of a SARS-CoV-2 protein other than the spike protein; the antigen comprises at least a portion of SARS-CoV-2 N protein (claim 9); and
A composition comprising a sensitizer that generates singlet oxygen in its excited state and is bound to a biotin-specific binding partner, directly or indirectly; the sensitizer is a photosensitizer (claim 13); the biotin-specific binding partner is avidin, an analog of avidin, and an antibody against biotin (claim 13).
The elements of a)-e) are in the same compartment (claim 11) or are split between two or more compartments (claim 12).
Claims 29-31 and 33-36
Claim 29 is directed to a microfluidics device comprising an inlet channel through which the samples is applied, and at least a first compartment capable of being in fluid communication with the inlet channel and containing:
A composition comprising:
A singlet oxygen-activatable chemiluminescent compound bound to a first target antigen, directly or indirectly; the antigen comprises an RBD of S1 protein (claim 30); and
A fluorescent molecule excited by the compound;
A composition comprising:
A singlet oxygen-activatable chemiluminescent compound bound to a second target antigen, directly or indirectly; wherein the antigen is at least a portion of a SARS-CoV-2 N protein (claim 31); and
A fluorescent molecule that is excited by the compound, wherein the fluorescent molecule is different from the fluorescent molecule of (a) and emits light at a different wavelength than the fluorescent molecule of (a);
At least one biotinylated anti-human Ig antibody; and
A composition comprising a sensitizer that generates singlet oxygen in its excited state and is bound to a biotin-specific binding partner, directly or indirectly; the sensitizer is a photosensitizer (claim 35); the biotin-specific binding partner is avidin, an analog of avidin, and an antibody against biotin (claim 35).
The elements of a)-e) are in the same compartment (claim 33) or are split between two or more compartments (claim 34).
The intended use of the microfluidic devices of claims for determining the presence and/or concentrations of multiple anti-SARS-CoV-2 antibodies in a sample. The singlet oxygen-activatable chemiluminescent compound of each of (a) and (b) is a substance that undergoes a chemical reaction with singlet oxygen to form a metastable intermediate species that can decompose with the simultaneous or subsequent emission of light (claims 13 and 35). The fluorescent molecules of (a) and (b) are each independently selected from terbium, uranium, samarium, europium, gadolinium and dysprosium (claims 14 and 36).
Claims 15-19, 21 and 22
Claim 15 is directed to a method for detecting the presence and/or concentration of multiple anti-SARS-CoV-2 antibodies in a sample. The method comprises the steps of:
Combining, either simultaneously or wholly or partially sequentially, a sample suspected of SARS-CoV-2 antibodies with a composition as outlined in claims 1-3, respectively (claims 18 and 19);
Allowing:
The binding of (a) and (c) to anti-SARS-CoV-2 spike protein antibodies in the sample;
The binding of (b) and (d) to anti-SARS-CoV-2 antibodies against the second target antigen in the sample; and
The binding of (c) and (d) to (e);
wherein the indirect binding of (a) to (d) results in the formation of a spike protein complex, and wherein the indirect binding of (b) to (e) results in the formation of a second target antigen complex, wherein in each of the complexes the sensitizer is brough into closer proximity to the chemiluminescent compound; and
Activating the sensitizer to generate singlet oxygen, wherein activation of the sensitizers present in the complexes causes activation of the chemiluminescent compound present in each complex;
Determining the amount of chemiluminescence generated by the activated compound in the spike protein complex by measuring the amount of light emitted by the fluorescent molecule of (a), wherein the amount of anti-SARS-CoV-2 spike antibodies in the sample is proportional to the amount of light emitted; and
Determining the amount of chemiluminescence generated by the activated compound in the second target antigen complex by measuring the amount of light emitted by the fluorescent molecule of (b), wherein the amount of anti-SARS-CoV-2 second target antigen antibodies in the sample is proportional to the amount of light emitted;
(claim 16) Determining that anti-SARS-CoV-2 antibodies present in the sample were generated in response to vaccination based on the result in step (4), and determining that anti-SARS-CoV-2 antibodies present in the sample were generated in response to infection based on the results of step (5). Steps (2)-(5) are repeated (claim 17).
Claims 37-41, 43 and 44
Claim 37 is directed to a method for detecting the presence and/or concentration of multiple anti-SARS-CoV-2 antibodies in a sample. The method comprises the steps of:
Combining, either simultaneously or wholly or partially sequentially, a sample suspected of SARS-CoV-2 antibodies with a composition as outlined in claims 23-25, respectively (claims 40 and 41);
Allowing:
The binding of (a) and (c) to anti-SARS-CoV-2 spike protein antibodies in the sample;
The binding of (b) and (c) to anti-SARS-CoV-2 antibodies against the second target antigen in the sample; and
The binding of (c) to (d);
wherein the indirect binding of (a) to (d) results in the formation of a spike protein complex, and wherein the indirect binding of (b) to (d) results in the formation of a second target antigen complex, wherein in each of the complexes the sensitizer is brough into closer proximity to the chemiluminescent compound; and
Activating the sensitizer to generate singlet oxygen, wherein activation of the sensitizers present in the complexes causes activation of the chemiluminescent compound present in each complex;
Determining the amount of chemiluminescence generated by the activated compound in the spike protein complex by measuring the amount of light emitted by the fluorescent molecule of (a), wherein the amount of anti-SARS-CoV-2 spike antibodies in the sample is proportional to the amount of light emitted; and
Determining the amount of chemiluminescence generated by the activated compound in the second target antigen complex by measuring the amount of light emitted by the fluorescent molecule of (b), wherein the amount of anti-SARS-CoV-2 second target antigen antibodies in the sample is proportional to the amount of light emitted;
(claim 38) Determining that anti-SARS-CoV-2 antibodies present in the sample were generated in response to vaccination based on the result in step (4), and determining that anti-SARS-CoV-2 antibodies present in the sample were generated in response to infection based on the results of step (5). Steps (2)-(5) are repeated (claim 39).
In the methods of claims 15-19, 21, 22, 37-41, 43 and 44, the singlet oxygen-activatable chemiluminescent compound of each of (a) and (b) is a substance that undergoes a chemical reaction with singlet oxygen to form a metastable intermediate species that can decompose with the simultaneous or subsequent emission of light (claims 21 and 43). The sensitizer is a photosensitizer, wherein the activation of the sensitizer in step (3) comprises irradiation with light (claims 21 and 43). The biotin-specific binding partner is avidin, an analog of avidin, and an antibody against biotin (claims 21 and 43). The biological sample is selected from whole blood, or any portion thereof, among other choices in claims 22 and 44.
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 1-3, 5-9, 11-19, 21-25, 27-31, 33-41, 43 and 44 are rejected under 35 U.S.C. 103 as being unpatentable over Burbelo et al. (JID, July 15, 2020, published online May 19, 2020, 222:206-213, “Burbelo”) in view of Ledden and Janzen (US 2016/0025736 A1, “Ledden”, cited in the IDS filed 12/17/2025), Wei (WO 2020/068548 A1, “Wei”), and Krishnamurthy et al. (November 19, 2020, PLoS ONE, 15(11):e0242655, 12 pages, “Krishnamurthy”).
Burbelo discloses quantitative measurements of plasma and serum antibodies to SARS-CoV-2 N and S proteins using a luciferase immunoprecipitation system (see abstract). Burbelo concludes that antibodies to the N protein are more sensitive for detecting early infection, in contrast with antibodies to the S protein, the target of vaccination, which are produced later (see page 206, right column, the full paragraph).
Burbelo’s method does not use the methods steps instantly claimed. However, it would have been obvious to have used the methods of Ledden and Wei (both Applicant’s own work) to accomplish the same objective as Burbelo. Ledden discloses chemiluminescent detection systems, kits and microfluid devices for medical diagnostics using LOCI technology, including the option to multiplex, with any target analyte (see Ledden, abstract and paragraph [0055]), and Wei discloses the same in a multiplex format for biotin (see Wei, abstract). Ledden’s and Wei’s methodologies, kits and microfluidic devices (see all claims of Ledden and Wei) align with the limitations of instant claims 1-3, 5-9, 11-19, 21-25, 27-31, 33-41, 43 and 44, as outlined above in the Claims Summary section. One would have been motivated to use the methodologies of Ledden and Wei, modified with the N and S antigens, to detect antibodies to the N and S proteins of SARS-CoV-2, since the methods are highly sensitive, allow for multiplexing, and do not require plasma separation (see Ledden, paragraph [0003]). One would have had a reasonable expectation of success given that chemiluminescent multiplex assays are used to detect antibodies to SARS-CoV-2 RBD and N proteins (see Krishnamurthy, page 1). Further, one would have been motivated to use the RBD, as is done by Krishnamurthy, since it is part of the S protein and considered an antigen is significance by Krishnamurthy (see page 1, “Methods”, and Tables 2-4). Therefore, the claimed embodiments would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention.
Conclusion
No claim is allowed.
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/STACY B CHEN/Primary Examiner, Art Unit 1672
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