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Heterologous radioimmunoassays for monkey gonadotrophins. I. Assessment of the reagents proposed for the assay of FSH

Journal Article:

Abstract

The reliability of the reagents used in heterologous immunoassay procedures for FSH in monkey sera was assessed. Re-investigation of the iodination and purification of the human FSH tracer confirmed the presence of different iodinated molecular species, such as a high molecular weight aggregated material, 'intact' human FSH and subunits. Tracers obtained with the chloramine T procedure were superior to those obtained by the lactoperoxidase method. Purification by cellulose adsorption chromatography resulted in partial dissociation of the intact hormone into free subunits, while Sephadex G-25 chromatography did not yield any subunit-like material. Purification by Ultrogel AcA 54 gave the best separation of the 'intact' iodinated FSH. Binding studies indicated that an anti-ovine FSH (oFSH) serum showed a high degree of binding to 'intact' hFSH, although its binding to subunits and to aggregated material was also significant. The highest degree of binding to this antiserum was shown by the 'intact' tracer iodinated with chloramine T and purified sequentially by Sephadex G-25 and Ultrogel (AcA 54) chromatography. Using a highly purified hFSH tracer in combination with three antisera (anti-oFSH, H 31; anti-hFSH, WHO, M-93-2; anti-hFSH, NIH, batch No. 5) displacement curves were investigated with three purified monkey FSH preparations. No displacement was observed  More>>
Authors:
Khan, S A; Diczfalusy, E [1] 
  1. Reproductive Endocrinology Research Unit, Karolinska sjukhuset, Stockholm
Publication Date:
Jan 01, 1983
Product Type:
Journal Article
Reference Number:
AIX-14-806199; EDB-84-031565
Resource Relation:
Journal Name: Acta Endocrinol.; (Denmark); Journal Volume: 104
Subject:
62 RADIOLOGY AND NUCLEAR MEDICINE; FSH; RADIOIMMUNOASSAY; MONKEYS; REAGENTS; RELIABILITY; ANIMALS; GONADOTROPINS; HORMONES; ISOTOPE APPLICATIONS; MAMMALS; PEPTIDE HORMONES; PITUITARY HORMONES; PRIMATES; RADIOASSAY; TRACER TECHNIQUES; VERTEBRATES; 550601* - Medicine- Unsealed Radionuclides in Diagnostics
OSTI ID:
5351496
Country of Origin:
Denmark
Language:
English
Other Identifying Numbers:
Journal ID: CODEN: ACENA
Submitting Site:
HEDB
Size:
Pages: 15-22
Announcement Date:

Journal Article:

Citation Formats

Khan, S A, and Diczfalusy, E. Heterologous radioimmunoassays for monkey gonadotrophins. I. Assessment of the reagents proposed for the assay of FSH. Denmark: N. p., 1983. Web.
Khan, S A, & Diczfalusy, E. Heterologous radioimmunoassays for monkey gonadotrophins. I. Assessment of the reagents proposed for the assay of FSH. Denmark.
Khan, S A, and Diczfalusy, E. 1983. "Heterologous radioimmunoassays for monkey gonadotrophins. I. Assessment of the reagents proposed for the assay of FSH." Denmark.
@misc{etde_5351496,
title = {Heterologous radioimmunoassays for monkey gonadotrophins. I. Assessment of the reagents proposed for the assay of FSH}
author = {Khan, S A, and Diczfalusy, E}
abstractNote = {The reliability of the reagents used in heterologous immunoassay procedures for FSH in monkey sera was assessed. Re-investigation of the iodination and purification of the human FSH tracer confirmed the presence of different iodinated molecular species, such as a high molecular weight aggregated material, 'intact' human FSH and subunits. Tracers obtained with the chloramine T procedure were superior to those obtained by the lactoperoxidase method. Purification by cellulose adsorption chromatography resulted in partial dissociation of the intact hormone into free subunits, while Sephadex G-25 chromatography did not yield any subunit-like material. Purification by Ultrogel AcA 54 gave the best separation of the 'intact' iodinated FSH. Binding studies indicated that an anti-ovine FSH (oFSH) serum showed a high degree of binding to 'intact' hFSH, although its binding to subunits and to aggregated material was also significant. The highest degree of binding to this antiserum was shown by the 'intact' tracer iodinated with chloramine T and purified sequentially by Sephadex G-25 and Ultrogel (AcA 54) chromatography. Using a highly purified hFSH tracer in combination with three antisera (anti-oFSH, H 31; anti-hFSH, WHO, M-93-2; anti-hFSH, NIH, batch No. 5) displacement curves were investigated with three purified monkey FSH preparations. No displacement was observed with a widely available anti-hFSH (WHO, M 93-2) serum. However, linear dose dependent displacement was found with another anti-hFSH (NIH, batch No. 5) serum and with an anti-oFSH (H 31) serum. Comparison of immunoassay with the human and ovine antiserum revealed significant differences in slope, parallelism, precision and effective range. Significant discrepancies in the FSH potency estimates in monkey sera were also observed with the two assay systems. It is suggested that - until a homologous system is developed - the immunoassay employing an hFSH tracer and an anti-oFSH serum should be used for macaque FSH.}
journal = {Acta Endocrinol.; (Denmark)}
volume = {104}
journal type = {AC}
place = {Denmark}
year = {1983}
month = {Jan}
}