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J Immunoassay Immunochem ; 34(4): 414-37, 2013.
Article in English | MEDLINE | ID: mdl-23859792

ABSTRACT

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.


Subject(s)
Antibodies, Monoclonal/immunology , Antigens/isolation & purification , Enzyme-Linked Immunosorbent Assay/methods , Hepatitis B Surface Antigens/immunology , Adsorption , Animals , Antibodies, Monoclonal/analysis , Antigen-Antibody Reactions , Buffers , Enzyme-Linked Immunosorbent Assay/instrumentation , Humans , Hydrogen-Ion Concentration , Immunoassay/methods , Rabbits
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