Immunogenicity assay
Inventors
Yin, Ray • Pan, Jing • Vallejo, Yli Remo • Small, Thomas • Qin, Dujie • Chen, De
Assignees
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Publication Number
US-10274501-B2
Publication Date
2019-04-30
Expiration Date
Abstract
Assays for detecting antibodies to pharmaceutical preparations, food allergens and environmental allergens are described.
Core Innovation
The described invention relates to immunochromatographic assays for detecting antibodies to an antigen, including a drug of interest, a food antigen, an environmental antigen, or a tag attached to the antigen. A lateral flow assay device includes a solid phase with an immobilized capture agent that comprises the antigen and a cognate tag of a member of a first binding pair, and reporter detection at the capture site correlates with presence of the antibody in the sample.
The invention uses a capture site configuration in which the solid phase comprises a branched polymer along with the immobilized capture agent, and it employs dual binding in which antigen comprising a member of a second binding pair is contacted after the initial capture. The antigen of the first capture interaction and the antigen contacted in the second interaction are the same, and a reporter is then contacted that comprises a cognate tag of the second binding pair and optionally comprises a branched polymer.
The document emphasizes improved detection performance for low-affinity/low-avidity antibodies by eliminating sample dilution and wash steps relative to conventional approaches. It further addresses high-dose hook effect concerns by using branched or dendritic polymers, including star, comb, dendrimer, and randomly branched polymers, to increase epitope availability.
Claims Coverage
The independent claim defines a complete method and recites the core inventive elements as capture on a solid phase with immobilized antigen and cognate tags using dual binding and reporter detection at the capture site, with the solid phase and reporter optionally involving branched polymers. Dependent claims further refine the antigen/tag, binding-pair sets, branched polymer types, and reporter types by adding enumerated selections as additional inventive limitations.
Antigen with cognate tag on immobilized capture agent and branched polymer solid phase
A method in which the capture site comprises a solid phase with an immobilized capture agent, where the solid phase comprises a member of a first binding pair and a branched polymer, and the capture agent comprises the antigen and a cognate tag of the member of the first binding pair.
Dual binding using same antigen for first and second binding pair
After contacting the sample to the capture site, contacting an antigen comprising a member of a second binding pair to the capture site, wherein the antigen of the capture agent interaction and the antigen of the second binding interaction are the same.
Reporter with cognate tag of second binding pair, optionally with branched polymer
Following dual binding, contacting a reporter to the capture site, where the reporter comprises a cognate tag of the member of the second binding pair and optionally a branched polymer.
Detect reporter at capture site correlating with antibody presence
Detecting the reporter at the capture site, wherein reporter at the capture site correlates with presence of the antibody to the antigen in the sample.
Drug of interest, food antigen, environmental antigen, or attached tag selection
The target antigen is selected as a drug of interest, a food antigen, an environmental antigen, or a tag attached to the drug of interest, food antigen, or environmental antigen.
PEG or PEO tag selection
The tag attached to the drug of interest, food antigen, or environmental antigen includes polyethylene glycol or polyethyleneoxide.
Enumerated first and second binding pair systems
The first and/or second binding pairs are selected from enumerated binding-system combinations including avidin/streptavidin/neutravidin with biotin; DNP with anti-DNP; digoxin with anti-digoxin; digoxigenin with anti-digoxigenin; hapten with anti-hapten antibodies; polysaccharide with polysaccharide binding moieties; lectin/receptor or ligand/receptor pairs; fluorescein with anti-fluorescein antibodies; and complementary nucleic acids.
Enumerated branched polymer types
The branched polymer is chosen from star-shaped polymer, comb-shaped polymer, dendrimer, starburst dendrimer, combburst dendrigraft, and hypercombbranched polymer.
Enumerated reporter types
The reporter is chosen from enzyme, ferritin, fluorescent or colored microparticle, bead, nanoparticle, colloid metal, quantum dot, magnetic particle, up-converting phosphorescent particle, electrochemiluminescent molecule, a compound comprising a transition metal, and a compound comprising a lanthanide metal.
Across the independent method and its listed dependents, the claim set centers on a dual binding immunochromatographic detection scheme in which antigen and cognate tags drive sequential binding, reporter detection at the capture site signals antibody presence, and branched polymers are used as part of the capture-site solid phase and optionally the reporter. Dependent claims further specify tag choices, binding-pair systems, branched polymer form factors, and labeled reporter types.
Stated Advantages
Improved detection of low-affinity/low-avidity antibodies by eliminating sample dilution and wash steps.
Mitigation of high-dose hook effect concerns by using branched/dendritic polymers to increase epitope availability.
Documented Applications
Immunochromatographic lateral flow assays for detecting anti-drug, anti-food-allergen, and anti-environmental-allergen antibodies across multiple formats, including bridge, competitive, and direct serologic formats.
Immunogenicity assay implementations for antibody isotype testing including IgG, IgM, IgA, IgE, and IgD.
Validation of detection performance via drug-depletion/competition and retesting of human serum samples.
Example lateral flow assay targets include anti-mouse IgG, anti-PEG/PEO, anti-glucagon, and anti-GLP-1.
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