Development of heterologous enzyme linked immunosorbent assay for dehydroepiandrosterone in serum.
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.
Department of Reproductive Biomedicine, National Institute of Health and Family Welfare, Munirka, New Delhi, India. tgsrivastava@nihfw.org
Tulsidas G
TG
Shrivastav
Shail K
SK
Chaube
Kiran P
KP
Kariya
Sana
S
Bhanot
Rita
R
Singh
Dinesh
D
Kumar
D000276:Adjuvants, Immunologic; D000818:Animals; D002417:Cattle; D003687:Dehydroepiandrosterone / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D006735:Horseradish Peroxidase; D007124:Immunoenzyme Techniques; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine
D000276:Adjuvants, Immunologic; D012710:Serum Albumin, Bovine; D003687:Dehydroepiandrosterone; D006735:Horseradish Peroxidase