Heterologous enzyme linked immunosorbent assay for measurement of testosterone in serum.
In steroid enzyme immunoassay (EIA), homologous and heterologous combinations of enzyme conjugate with immunogen influences labeled steroid recognition by antibodies that affect sensitivity of the assay. To develop testosterone enzyme linked immunosorbent assay (ELISA), antibodies were generated against testosterone-3-carboxymethyloxime-bovine serum albumin (T-3-CMO-BSA), testosterone-11-hemisuccinate-bovine serum albumin (T-11-HS-BSA), testosterone-17-hemisuccinate-bovine serum albumin (T-17-HS-BSA), testosterone-17-glucuronide-bovine serum albumin (T-17-G-BSA), and testosterone-19-carboxymethylether-bovine serum albumin (T-19-CME-BSA). Testosterone horseradish peroxidase (HRP) enzyme conjugate were prepared using carboxyl derivatives of 11-keto-testosterone (11-keto-T) and 1-dehydrotestosterone (1-Dehydro-T). Ten combinations of heterologous assays were evaluated. The data of the present study revealed that the use of the T-11-HS-BSA antibody in antigen plus site heterologous assay that employed 11-keto-testosterone-3-CMO-HRP as the label showed binding and displacement, and led to the development of sensitive and specific 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
Pramod K V
PK
Prasad
Kiran P
KP
Kariya
Dinesh
D
Kumar
D000293:Adolescent; D000328:Adult; D000818:Animals; D000910:Antibodies, Heterophile / Q000276:immunology; D015478:Antigens, Heterophile / Q000097:blood* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006735:Horseradish Peroxidase / Q000737:chemistry; D006801:Humans; D008297:Male; D011817:Rabbits; D012710:Serum Albumin, Bovine / Q000276:immunology; D013739:Testosterone / Q000031:analogs & derivatives / Q000097:blood* / Q000276:immunology; D055815:Young Adult
D000910:Antibodies, Heterophile; D015478:Antigens, Heterophile; D012710:Serum Albumin, Bovine; D013739:Testosterone; D006735:Horseradish Peroxidase