Production and characterization of an antiserum to synthetic gonadotropin-releasing hormone |
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Authors: | Y Koch M Wilchek M Fridkin P Chobsieng U Zor H R Lindner |
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Institution: | 4. Department of Biodynamics The Weizmann Institute of Science, Rehovot, Israel;1. Department of biophysics The Weizmann Institute of Science, Rehovot, Israel;2. Department of Chemistry The Weizmann Institute of Science, Rehovot, Israel |
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Abstract: | The synthetic decapeptide “luteinizing hormone-releasing hormone” (LH-RH) was rendered antigenic by reaction of its histidine or tyrosine residues (7 : 3 approx.) with p-diazonium phenylacetic acid and coupling of the azo-derivatives formed to bovine serum albumin (BSA). Immunization of rabbits yielded antisera that bound 125I-labeled LH-RH (approx. 50 pg) at dilutions up to 1:200, 000 and showed no cross-reaction with unrelated hypothalamic and pituitary hormones, extracts from rat cerebral cortex, and with small fragments of LH-RH. Cross-reaction was minimal (0.2%) with the free acid analogue of LH-RH, and moderate with -pGlu LH-RH (20%), -pGlu-His-LH-RH (2.4%) and with LH-RH analogues in which a single residue (No. 4–6 or No. 8) was exchanged by an amino-acid of similar character (1.2–12%). Biologically active hypothalamic extract and LH-RH produced parallel 125I-LH-RH-binding inhibition curves, providing immunochemical support for the identity of the native releasing hormone with synthetic LH-RH. |
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