The antigenic determinant of the LH-releasing hormone for three different antisera.

A. Arimura, H. Sato, D. H. Coy, R. B. Worobec, A. V. Schally, N. Yanaihara, T. Hashimoto, C. Yanaihara, N. Sukura

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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-reactivity. Histidine in position 2 played an important role but neither the amide group nor the glycine amide group at the C-terminus were essential. For antiserum No. 742, the C-terminal tetrapeptide-amide fragment of LH-RH showed considerable cross-reactivity in the LH-RH, the amide moiety itself being of crucial importance. These antisera may be useful in investigating peptides related to LH-RH in biological materials.

Original languageEnglish (US)
Pages (from-to)222-231
Number of pages10
JournalActa endocrinologica
Volume78
Issue number2
DOIs
StatePublished - Feb 1975

ASJC Scopus subject areas

  • Endocrinology

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    Arimura, A., Sato, H., Coy, D. H., Worobec, R. B., Schally, A. V., Yanaihara, N., Hashimoto, T., Yanaihara, C., & Sukura, N. (1975). The antigenic determinant of the LH-releasing hormone for three different antisera. Acta endocrinologica, 78(2), 222-231. https://doi.org/10.1530/acta.0.0780222