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Preparation of specific antisera to 15alpha-hydroxyestrogen 15-N-acetylglucosaminides.
Authors:M Nakagomi  S Iida  Y Hara  Y Matsuki  T Nambara  E Suzuki
Affiliation:Hatano Research Institute, Food and Drug Safety Center, Hadano, Kanagawa, Japan. madokan@qc4.so-net.ne.jp
Abstract:3-(1-Carboxypropyl) ether derivatives of 15alpha-hydroxyestradiol 15-N-acetylglucosaminide (15alpha-OHE2 15NAG) and 15alpha-hydroxyestriol (E4) 15NAG were synthesized and conjugated with bovine serum albumin. Antisera elicited in rabbits possessed high affinity and specificity for the 15alpha-hydroxyestrogen (15alpha-OHEs) 15NAG, exhibiting no significant cross-reactivity with 15alpha-OHEs and their positional isomers such as 16NAG and 17NAG. Enzyme immunoassay methods developed by using the purified antisera and horseradish peroxidase-labeled antigens were applied to the measurement of 15alpha-OHEs 15NAG and E4 15NAG in normal pregnancy urine. We demonstrated for the first time that the conjugation of N-acetylglucosamine to E4 occurs at the C-15alpha position.
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