title: Antigen heterologous enzyme linked immunosorbent assay for the measurement of DHEA in serum.
abstract: Homologous and heterologous combinations of enzyme conjugate and antibody in steroid enzyme immunoassay (EIA) influences unlabeled steroid recognition by antibody that affects sensitivity of the assay. To develop dehydroepiandrosterone (DHEA) antigen heterologous enzyme linked immunosorbent assay (ELISA), antibodies were generated against DHEA-3-hemisuccinate-bovine serum albumin (DHEA-3-HS-BSA), DHEA-7-carboxymethyloxime-bovine serum albumin (DHEA-7-CMO-BSA), and DHEA-17-carboxymethyloxime-bovine serum albumin (DHEA-17-CMO-BSA). Five horseradish peroxidase (HRP) enzyme conjugates were prepared using five testosterone derivatives [testosterone-3-CMO (T-3-CMO), testosterone-17-HS (T-17-HS), testosterone-17-glucuronoside (T-17-G), testosterone-19-carboxymethylether (T-19-CME), and testosterone-11-HS (T-11-HS)]. Fifteen antigen heterologous combinations of antibody and enzyme conjugates were evaluated in the standard binding assay; only two combinations showed binding. The use of antigen heterologous combination (different antigen in label than the immunogen) resulted in development of a simple, direct, and convenient assay as it permits the direct addition of the serum sample into the assay and it requires only 1.5 h to complete.
authors: Department of Reproductive Biomedicine, National Institute of Health and Family Welfare, Munirka, New Delhi, India. tgsrivastava@nihfw.org
authors: Tulsidas G
authors: TG
authors: Shrivastav
authors: Shail K
authors: SK
authors: Chaube
authors: Kiran P
authors: KP
authors: Kariya
authors: Dinesh
authors: D
authors: Kumar
authors: Rita
authors: R
authors: Singh
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D015478:Antigens, Heterophile; D002417:Cattle; D003687:Dehydroepiandrosterone / Q000031:analogs & derivatives / Q000032:analysis* / Q000097:blood / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism; D010091:Oximes / Q000032:analysis / Q000097:blood / Q000276:immunology; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000276:immunology; D013739:Testosterone / Q000031:analogs & derivatives / Q000032:analysis / Q000097:blood / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D015478:Antigens, Heterophile; D010091:Oximes; C106648:dehydroepiandrosterone-7-O-(carboxymethyl)oxime; C509625:dehydroepiandrosterone-bovine serum albumin; C067733:testosterone-3-carboxymethyloxime; D012710:Serum Albumin, Bovine; D013739:Testosterone; D003687:Dehydroepiandrosterone; C017109:19-O-carboxymethoxytestosterone; D006735:Horseradish Peroxidase