Preparation of specific antiserum to 17 alpha-estradiol 17-N-acetylglucosaminide.
A specific enzyme immunoassay (EIA) method has been developed for 17 alpha-estradiol 17-N-acetylglucosaminide as a model compound instead of 15 alpha-hydroxyestrogen 15-N-acetylglucosaminides. Two new haptens, 3-(omega-carboxyalkyl) ether derivatives of 17-alpha-estradiol 17-N-acetylglucosaminide, were synthesized and conjugated with bovine serum albumin (BSA). The EIA was newly established using specific antiserum elicited against 3-(1-carboxypropyl) ether of 17 alpha-estradiol 17-N-acetylglucosaminide (17NAG CPE)-BSA conjugate and beta-galactosidase-labeled 17NAG CPE as a labeled antigen. An appropriate dose-response curve of EIA for 17 alpha-estradiol 17-N-acetylglucosaminide was obtained in the range of 20-1000 pg/tube. The specificity of EIA proved to be satisfactory in terms of cross-reactivities to related compounds including 15 alpha-N-acetylglucosaminides. The proposed method will be applicable to the preparation of antisera for use in EIA of 15 alpha-hydroxyestrogen 15-N-acetylglucosaminides.
Suzuki
E
E
Hatano Research Institute, Food and Drug Safety Center, Kanagawa, Japan.
Nakagomi
M
M
Tsuruta
S
S
Yamawaki
C
C
Goto
J
J
Matsuki
Y
Y
Nambara
T
T
D000117:Acetylglucosamine / Q000032:analysis* / Q000276:immunology; D000818:Animals; D004958:Estradiol / Q000032:analysis* / Q000276:immunology; D007106:Immune Sera / Q000276:immunology*; D007124:Immunoenzyme Techniques; D008297:Male; D011817:Rabbits; D012710:Serum Albumin, Bovine
D007106:Immune Sera; D012710:Serum Albumin, Bovine; D004958:Estradiol; D000117:Acetylglucosamine