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Amplified Immunoassay of Human IgG Using Real‐time Biomolecular Interaction Analysis (BIA) Technology
Author(s) -
Pei RenJun,
Cui XiaoQiang,
Yang XiuRong,
Wang ErKang
Publication year - 2002
Publication title -
chinese journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.28
H-Index - 41
eISSN - 1614-7065
pISSN - 1001-604X
DOI - 10.1002/cjoc.20020200507
Subject(s) - chemistry , surface plasmon resonance , polyclonal antibodies , detection limit , immunoassay , covalent bond , antibody , chromatography , immunoglobulin g , biosensor , protein g , microbiology and biotechnology , nanoparticle , nanotechnology , biochemistry , immunology , materials science , organic chemistry , biology
An automated biomolecular interaction analysis instrument (BI‐Acore) based on surface plasmon resonance (SPR) has been used to determine human immunoglobulin G (IgG) in real time. Polyclonal and‐human IgG antibody was covalently immobilized to a carboxymethyldextran‐modified gold film surface. The samples of human IgG prepared in HBS buffer were poured over the immobilized surface. The signal amplification antibody was applied to amplify the response signal. After each measurement, the surface was regenerated with 0.1 mol/L H 3 PO 4 . The assay was rapid, requiring only 30 min for antibody immobilization and 20 min for each subsequent process of immune binding, antibody amplification and regeneration. The antibody immobilized surface had good response to human IgG in the range of 0.12–60 nmol/L with a detection limit of 60 pmol/L. The same antibody immobilized surface could be used for more than 110 cycles of binding, amplification and regeneration. The results demonstrate that the sensitivity, specificity and reproducibility of amplified immunoassay using real‐time BIA technology are satisfactory.