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Radioimmunoassay of Bovine, Ovine and Porcine Luteinizing Hormone with a Monoclonal Antibody and a Human Tracer

Acta Veterinaria Scandinavica volume 34, pages 255–262 (1993)Cite this article

Abstract

A radioimmunoassay for bovine (bLH), ovine (oLH) and porcine (pLH) luteinizing hormone was developed using a human 125 ILH tracer from a commercial kit and a monoclonal antibody (518B7) specific for LH but with low species specificity. Standard curves demonstrated similar binding kinetics when bLH, oLH and pLH were incubated with tracer and antibody for 2 h at room temperature. A 30-min delay in the addition of the tracer gave sufficient sensitivity when analysing pLH. Separation of antibody-bound LH from free hormone was achieved by using second antibody-coated micro Sepharose beads. The assay was validated and the performance compared with that of an RIA currently in use for determination of bLH and oLH (coefficient of correlation: 0.99 and 0.98). Regardless of the standards used, intra-assay coefficients of variation were <10% for LH concentrations exceeding 1 µg/L. The inter-assay coefficients of variation were <15%. The assay was used for clinical evaluation demonstrating the pre-ovulatory LH surge in two cyclic cows, LH pulsatility in an oophorectomized ewe and LH response to GnRH injection in a boar.

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Acknowledgments

We thank Dr. Janet Roser for supplying the LH anti-sera. Thanks are also due to the USDA Animal Hormone Program, the NIAMDD-NPA Pituitary Distribution Program, the NIADDK-NIH and Dr. Leo Reichert for supplying hormone preparations. The Swedish Agency for Research in Developing Countries (SAREC) and the Swedish Institute are acknowledged for financial support.

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Authors and Affiliations

  1. Department of Clinical Chemistry and Department of Physiology, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, P.O. Box 7038, S-750 07, Uppsala, Sweden

    M. Forsberg, R. Tagle, A. Mad, J.R. Molina & M.-A. Carlsson

  2. Faculty of Veterinary Medicine, Laboratory of Nuclear Technique, Las Places 1550, Montevideo, Uruguay

    M. Forsberg, R. Tagle, A. Mad, J.R. Molina & M.-A. Carlsson

  3. School of Agronomy, University of Costa Rica, San Jose’, Costa Rica

    M. Forsberg, R. Tagle, A. Mad, J.R. Molina & M.-A. Carlsson

Authors
  1. M. Forsberg
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  2. R. Tagle
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  3. A. Mad
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  4. J.R. Molina
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  5. M.-A. Carlsson
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Forsberg, M., Tagle, R., Mad, A. et al. Radioimmunoassay of Bovine, Ovine and Porcine Luteinizing Hormone with a Monoclonal Antibody and a Human Tracer. Acta Vet Scand 34, 255–262 (1993). https://doi.org/10.1186/BF03548189

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  • DOI: https://doi.org/10.1186/BF03548189

Keywords

Acta Veterinaria Scandinavica

ISSN: 1751-0147