DEVELOPMENT OF HETEROLOGOUS ENZYME LINKED IMMUNOSORBENT ASSAY FOR DEHYDROEPIANDROSTERONE IN SERUM

Abstract

Homologous and heterologous combinations of enzyme conjugate with immunogen in steroid enzyme immunoassay (EIA) influence labeled steroid recognition by an antibody that affects the sensitivity of the assay. To develop dehydroepiandrosterone (DHEA) enzyme linked immunosorbent assay (ELISA), antibodies were generated against dehydroepiandrosterone-17-carboxymethyloxime-bovine serum albumin (DHEA-17-CMO-BSA) and dehydroepiandrosterone-7-carboxymethyloxime- bovine serum albumin (DHEA-7-CMO-BSA). Two dehydroepiandrosterone (DHEA) horse radish peroxidise (HRP) enzyme conjugates were prepared using two dehydroepiandrosterone derivatives (DHEA-17-CMO and DHEA-7-CMO). Four combinations of homologous and heterologous assays were evaluated. The use of heterologous combination improved the sensitivity of the assay.

Keywords:

• dehydroepiandrosterone
• heterologous
• homologous
• immunoassay
• site heterology

ACKNOWLEDGMENTS

The National Institute of Health and Family Welfare, New Delhi, India, supported this study. The authors are grateful to Prof. D. Nandan and K. Kalaivani for their keen interest and encouragement in the present study.

Notes

n = Number of times same sample was analyzed for intraassay variation; N = number of times assays were carried out for interassay variation.