Effect of conditions of monoclonal antibody adsorption on antigen-binding activity

Autor:	O. V. Fartushnaya, D. A. Yakovleva, Yu. N. Tarakanova, Yu. S. Massino, G. I. Kolyaskina, Segal Ol, M. D. Smirnova, Dmitriev Ad, V. F. Lavrov, D. A. Dmitriev
Rok vydání:	2012
Předmět:	HBsAg Chromatography biology Chemistry medicine.drug_class Monoclonal antibody Applied Microbiology and Biotechnology Biochemistry Horseradish peroxidase Molecular biology Virus Adsorption Antigen Biotinylation biology.protein medicine Antibody
Zdroj:	Applied Biochemistry and Microbiology. 48:506-512
ISSN:	1608-3024 0003-6838
DOI:	10.1134/s0003683812050122
Popis:	The dependence of the antigen-binding activity of immobilized antibodies on pH of a saturating buffer has been investigated. We analyzed 28 monoclonal antibodies (MCAs) produced by various hybridomas to three virus antigens, i.e., the nuclear p23 protein of hepatitis C virus (C core protein p23), p24 protein of HIV 1, and the surface antigen of hepatitis B virus (HBsAg). Antibodies were adsorbed on the surfaces of immune plates in acidic (pH 2.8), neutral (pH 7.5), and alkaline (pH 9.5) buffers. The binding of labeled antigens, i.e., biotinylated or conjugated with horseradish peroxidase, with immobilized antigens was tested. It was shown that 10 out of 28 analyzed MCAs (36%) considerably better preserved their antigen-binding activity if their passive adsorption was carried out on the surface of polystyrene plates in an acidic buffer (pH 2.8). This approach allowed constructing a highly sensitive sandwich method for HBsAg assay with a minimal reliably determined antigen concentration of 0.013–0.017 ng/ml. The described approach may be recommended for the optimization of sandwich methods and solid-phase competitive methods.
Databáze:	OpenAIRE
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