Preparation of specific antiserum to 17 alpha-estradiol 17-N-acetylglucosaminide

Biol Pharm Bull. 1996 Nov;19(11):1396-400. doi: 10.1248/bpb.19.1396.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylglucosamine / analysis*
  • Acetylglucosamine / immunology
  • Animals
  • Estradiol / analysis*
  • Estradiol / immunology
  • Immune Sera / immunology*
  • Immunoenzyme Techniques
  • Male
  • Rabbits
  • Serum Albumin, Bovine

Substances

  • Immune Sera
  • Serum Albumin, Bovine
  • Estradiol
  • Acetylglucosamine