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Study on Antibody-enzyme Coupling and Enzyme Immunoassay Methods  

Jang Sean Il (Department of Skin & Beauty, Seojeong College)
Publication Information
Journal of Physiology & Pathology in Korean Medicine / v.18, no.3, 2004 , pp. 874-879 More about this Journal
Abstract
Alakaline phosphatase (ALP)- or horseradish peroxidase (HRP)-antibody conjugate was used frequently on the immunological detection methods such as enzyme-linked immunosobent assay (ELISA), immunobolt, immunohistochemistry. The classical enzyme-antibody coupling method by one-step (direction) injection of glutaraldehyde bring into being disadvantage such as low sensitivity of antigen detection because of homopolymers. This study was modified with the dialysis glutaraldehyde method to provide simple coupling through E-amino residues present in most protein. The dialysis glutaraldehyde coupling effects were better than the classical one-step glutaraldehyde injection in antigen detection of ELISA and immunobolt. Optimal dose of the dialysis glutaraldehyde solution was 0.10-0.25 %. This results suggest that the dialysis glutaraldehyde coupling method can readily applied to antigen detection of in vitro and in vivo.
Keywords
Alkaline phosphatase; horseradish peroxidase; dialysis; glutaraldehyde; antibody; coupling method;
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