# US 2022/0034896 A1 · Methods for quantitative analysis of one or more biomarkers > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2022/0034896 A1 - Document ID: US20220034896A1 - Status: Published application - Kind: A1 - Application number: 17/368,711 - Filed: 2021-07-06 - Published: 2022-02-03 - Earliest claimed: 2020-07-03 - Assignees: The Research Foundation for The State University of New York - Applicants: The Research Foundation for the State University of New York - Inventors: Nathaniel Cady - Web page: https://patentgrep.com/patent/US20220034896A1 ## CPC - G01N 33/54373: {involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings} - G01N 33/6854: {Immunoglobulins} - G01N 21/6428: {Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53)} - G01N 21/6452: {Individual samples arranged in a regular 2D-array, e.g. multiwell plates} - G01N 21/648: {using evanescent coupling or surface plasmon coupling for the excitation of fluorescence} - G01N 33/582: {with fluorescent label} - G01N 33/56983: {Viruses} - G01N 2333/165: Coronaviridae, e.g. avian infectious bronchitis virus - G01N 2800/26: Infectious diseases, e.g. generalised sepsis - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract The present disclosure relates to apparatuses and methods for detecting the amount and/or type of one or more analytes-of-interests such as biomarkers in a sample. In embodiments, the disclosure includes a method for determining a humoral response due to the presence of a target infectious agent or vaccine. In embodiments, detecting emission light from one or more fluorescent complexes is used to determine a type and/or quantity of the plurality of biomarkers. ## Claims ### Claim 1 A method for determining a humoral response due to a presence of a target infectious agent or vaccine, comprising: capturing one or more biomarkers on a substrate that binds a plurality of biomarkers to one or more binding sites, wherein one or more binding sites bind one or more biomarkers, and wherein when one or more biomarkers are present, forming one or more bound biomarkers-of-interest; contacting the one or more bound biomarkers-of-interest with one or more fluorescent binding partners to form one or more fluorescent complexes; contacting the substrate with a source of collimated, polarized light, wherein the source of collimated polarized light is configured to emit light at a predetermined wavelength appropriate for transferring energy to a plurality of surface plasmons and excite fluorescence of the one or more fluorescent complexes; detecting emission light of the one or more fluorescent complexes; and determining a type and/or quantity of the plurality of biomarkers. ### Claim 2 (depends on 1) The method of claim 1, wherein the one or more biomarkers are immunoglobulins. ### Claim 3 (depends on 2) The method of claim 2, wherein the immunoglobulins comprise one or more of immunoglobulin G (IgG), immunoglobulin M (IgM), immunoglobulin A (IgA), or isotypes or combinations thereof. ### Claim 4 (depends on 3) The method of claim 3, wherein the immunoglobulins comprise one or more of immunoglobulin G (IgG) directed against SARS-CoV-2, a SARS-CoV-2 variant, and/or a COVID 19 vaccine, immunoglobulin M (IgM) directed against SARS-CoV-2, a SARS-CoV-2 variant, and/or a COVID 19 vaccine, immunoglobulin A (IgA) directed against SARS-CoV-2, a SARS-CoV-2 variant, and/or a COVID 19 vaccine, or isotypes or combinations thereof. ### Claim 5 (depends on 1) The method of claim 1, wherein the one or more biomarkers are serum antibody targets. ### Claim 6 (depends on 1) The method of claim 1, wherein the substrate is disposed within a cell. ### Claim 7 (depends on 6) The method of claim 6, wherein the cell is a fluidic flow cell or a test-tube. ### Claim 8 (depends on 1) The method of claim 1, wherein each binding site of the plurality of binding sites comprises two or more predetermined antigens configured to bind to two or more different predetermined biomarkers-of-interest. ### Claim 9 (depends on 1) The method of claim 1, wherein the plurality of biomarkers are derived from blood serum, blood plasma, whole blood, dried blood. ### Claim 10 (depends on 1) The method of claim 1, wherein the type is characterized as immunoglobulin G (IgG), immunoglobulin M (IgM), immunoglobulin A (IgA), or isotypes or combinations thereof. ### Claim 11 (depends on 1) The method of claim 1, wherein the type is further characterized as virus specific to COVID-19, SARS-CoV-2, or a variant of SARS-CoV-2. ### Claim 12 (depends on 1) The method of claim 1, wherein the plurality of biomarkers are antibodies specific to a SARS-COV-2 full-length spike protein (SEQ ID NO:1), SARS-COV-2 spike 1 protein (SEQ ID NO: 2), SARS-COV-2 envelope protein (N) (SEQ ID NO: 3). ### Claim 13 (depends on 1) The method of claim 1, wherein contacting the substrate with a source of collimated, polarized light, further comprises positioning the substrate within a grating-coupled fluorescent plasmonic (GC-FP) detection platform. ### Claim 14 (depends on 1) The method of claim 1, wherein the source of collimated polarized light is configured to cause fluorescent emission intensity greater than 10×, greater than 100×, greater than 500×, or greater than 1000×. ### Claim 15 (depends on 1) The method of claim 1, wherein detecting emission light of the one or more fluorescent complexes further comprises forming a fluorescent image on an antigen array or map of antigens upon the substrate. ### Claim 16 (depends on 1) The method of claim 1, wherein detecting emission light of the one or more fluorescent complexes further comprising normalizing a fluorescence intensity of a plurality of spots to a control spot. ### Claim 17 (depends on 16) The method of claim 16, wherein normalizing further comprises generating a GC-FP detection ratio. ### Claim 18 An apparatus for quantitative analysis of a plurality of biomarkers contained in a sample, comprising: a substrate comprising a plurality of binding sites, one or more of the plurality of binding sites comprises one or more predetermined antigens configured to bind to one or more predetermined biomarkers-of-interest and form one or more bound biomarkers-of-interest when biomarkers-of-interest are flowed over the substrate, and wherein the biomarkers-of-interest, when present, are bind one or more fluorescent binding partners to form one or more fluorescent complexes; and a grating-coupled fluorescent plasmonic (GC-FP) detection platform, wherein the platform contacts the substrate with a source of collimated, polarized light, wherein the source of collimated polarized light emits light at a predetermined wavelength appropriate for transferring energy to a plurality of surface plasmons and excite fluorescence of the one or more fluorescent complexes, wherein the one or more biomarkers-of-interest are selected from immunoglobulins comprising one or more of immunoglobulin G (IgG) directed against SARS-CoV-2, a SARS-CoV-2 variant, and/or a COVID 19 vaccine, immunoglobulin M (IgM) directed against SARS-CoV-2, a SARS-CoV-2 variant, and/or a COVID 19 vaccine, immunoglobulin A (IgA) directed against SARS-CoV-2, a SARS-CoV-2 variant, and/or a COVID 19 vaccine, or isotypes or combinations thereof. ### Claim 19 (depends on 18) The apparatus of claim 18, wherein the apparatus detects and normalizes fluorescence of the one or more fluorescent complexes. ### Claim 20 A method for determining a humoral response due to a presence of a target infectious agent or vaccine, comprising: detecting in a biological sample of a subject at least one target antibody that is susceptible of being produced by the subject when the subject is vaccinated or infected by a target infectious agent, wherein the method comprises: capturing the at least one target antibody on a substrate that binds the at least one target antibody to a plurality of binding sites, wherein one or more binding sites bind one or more target antibodies, and wherein when one or more target antibodies are present forming one or more bound biomarkers-of-interest; contacting the one or more bound biomarkers-of-interest with one or more fluorescent binding partners to form one or more fluorescent complexes; contacting the substrate with a source of collimated, polarized light, wherein the source of collimated polarized light emits light at a predetermined wavelength appropriate for transferring energy to a plurality of surface plasmons and excite fluorescence of the one or more fluorescent complexes; detecting emission light of the one or more fluorescent complexes; and determining a type and/or quantity of the at least one target antibody, wherein the at least one target antibody is selected from immunoglobulins comprising one or more of immunoglobulin G (IgG) directed against SARS-CoV-2, a SARS-CoV-2 variant, and/or a COVID 19 vaccine, immunoglobulin M (IgM) directed against SARS-CoV-2, a SARS-CoV-2 variant, and/or a COVID 19 vaccine, immunoglobulin A (IgA) directed against SARS-CoV-2, a SARS-CoV-2 variant, and/or a COVID 19 vaccine, or isotypes or combinations thereof.