The Antigenic Determinant of the Lh-Releasing Hormone for Three Different Antisera

Abstract

Antigenic determinants of LH-releasing hormone (LH-RH) were investigated by testing the cross-reaction of LH-RH analogues and fragments in LH-RH radioimmunoassay (RIA) systems using 3 different antisera against the LH-RH decapeptide. Rabbit antiserum No. 419 was generated against LH-RH adsorbed on polyvinylpyrrolidone (PVP). Antisera Nos. 710 and 742 were produced by immunizing rabbits with LH-RH conjugated either with bovine serum albumin through its C-terminus, or with human serum albumin through the N-terminus, respectively. For antiserum No. 419, the N-terminal (pyro)-glutamic acid and/or histidine in positions 1 and 2 of LH-RH, respectively, were found to enhance the antigen-antibody interaction, but were not indispensable for it. Similarly, the C-terminal amide and glycine-NH2 did not play a major role in these interactions. The LH-RH heptapeptide fragment, corresponding to amino acid sequence from positions 3 to 9, showed a cross-reactivity in this RIA system with LH-RH, although greater amounts than those of cold LH-RH were required for a comparable inhibition of binding of labelled LH-RH. For antiserum No. 710, the LH-RH hexapeptide fragment corresponding to positions 2 to 7 showed considerable cross-reac-

Immunohistochemical Demonstration of proGnRH and GnRH in the Preoptic-Basal Hypothalamus of the Primate

“…The 6-16 fragment was conjugated to thyroglobulin using the carbodiimide method [2]. The resulting solution was diluted in sa line and mixed with Freund's complete adjuvant for immunization of 3 young adult female rabbits [3].…”

Preparation of a Monoclonal Antibody to Common Amino Acid Sequence of LHRH and Its Application

“…These results revealed the molecular multiplicity of LHRH. Many kinds of polyclonal anti-LHRH sera have been prepared and contributed to the studies on LHRH (Arimura et al, 1973(Arimura et al, , 1975Copeland et al, 1979), however, they could be used only for limited species because of strict species specificity, and also could not detect possible precursors because of the special structure of LHRH in the Nand C-terminals. Recently, monoclonal antibodies which were prepared by hybridoma technology (Kohler and Milstein, 1975) with highly specific binding sites, have been used widely as immunochemical probes.…”

The Effect of Ovarian Steroid Feedback upon Radioimmunoreactive Luteinizing Hormone Releasing Hormone in the Hypothalamus

“…Arimura et al(1975) (Harris, 1955;Kobayashi, 1956). Our data demonstrated an apparent increase of the radioimmunoreactive LH-RH in the hypothalamus 7 days after ovariectomy.…”