MONOCLONAL ANTIBODIES REQUIRING COATING BUFFER WITH LOW PH FOR EFFICIENT ANTIGEN CAPTURE IN SANDWICH ELISA: THE RARITIES OR PRACTICALLY IMPORTANT PHENOMENA?

This article reexamines some opinions concerning pH requirements for optimal immobilization of monoclonal antibodies (mAbs) by passive adsorption in antigen capture ELISA. It was discovered that substitution of "classical" sodium phosphate (pH 7.5) and carbonate (pH 9.5) coating solutions...

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Bibliographic Details
Published in:Journal of immunoassay & immunochemistry 2013-10, Vol.34 (4), p.414-437
Main Authors: Dmitriev, Alexander D., Tarakanova, Julia N., Yakovleva, Dinora A., Dmitriev, Dmitriy A., Phartooshnaya, Olga V., Kolyaskina, Galina I., Massino, Yulia S., Borisova, Olga V., Segal, Olga L., Smirnova, Maria B., Ulanova, Tatiana I., Lavrov, Viacheslav F.
Format: Article
Language:English
Subjects:
Adsorption
analytical sensitivity
Animals
Antibodies, Monoclonal - analysis
Antibodies, Monoclonal - immunology
antibody adsorption
Antigen-Antibody Reactions
Antigens - isolation & purification
Buffers
coating buffer pH
Enzyme-Linked Immunosorbent Assay - instrumentation
Enzyme-Linked Immunosorbent Assay - methods
Hepatitis B Surface Antigens - immunology
Humans
Hydrogen-Ion Concentration
Immunoassay - methods
immunoassay optimization
monoclonal antibodies
Rabbits
sandwich ELISA
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Summary:This article reexamines some opinions concerning pH requirements for optimal immobilization of monoclonal antibodies (mAbs) by passive adsorption in antigen capture ELISA. It was discovered that substitution of "classical" sodium phosphate (pH 7.5) and carbonate (pH 9.5) coating solutions by acid (pH 2.8) buffers maximized antigen capture 4 out of 10 different tested anti-HBsAg mAbs, resulting in a 1.5-2.5 increase of binding curve coefficients. By measuring both mAbs amounts and functionality, the enhancement effect was attributed to the better preservation of solid phase antibodies activity.
ISSN:1532-1819
1532-4230
DOI:10.1080/15321819.2013.764894