1. Noncompetitive Fluorescence Polarization Immunoassay for Protein Determination
- Author
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Koji Shigemura, Ayuko Imai, Akihide Hibara, Keine Nishiyama, Ayano Nakamura, Manabu Tokeshi, and Mao Fukuyama
- Subjects
Detection limit ,Heavy chain ,Chromatography ,medicine.diagnostic_test ,biology ,Chemistry ,Fluorescent polarization ,Single-Domain Antibodies ,Immunoglobulin G ,Analytical Chemistry ,Immunoassay method ,Limit of Detection ,Immunoassay ,Fluorescence Polarization Immunoassay ,Fluorescence polarization immunoassay ,medicine ,biology.protein ,Animals ,Humans ,Rabbits ,Antibody - Abstract
Fluorescent polarization immunoassay (FPIA) is a single-step immunoassay method that is applicable to point-of-care testing; however, its applicability to large biomolecules has been restricted because ordinary FPIA is a competitive assay. Here, we report a noncompetitive FPIA using the variable domain from the heavy chain of a camelid antibody (VHH antibody). FPIA with VHH was successfully used to quantitate rabbit immunoglobulin G (IgG) and demonstrated a wider response range than that observed with antibody-binding (Fab) fragment. Then, using a portable FPIA instrument, a VHH-based immunoassay of human IgG in a human serum certified reference material was demonstrated.
- Published
- 2020
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