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Related Experiment Videos

Circulating plasma corticotrophin-releasing factor-like immunoreactivity.

M J Ellis1, J H Livesey, R A Donald

  • 1Department of Endocrinology, Princess Margaret Hospital, Christchurch, New Zealand.

The Journal of Endocrinology
|May 1, 1988
PubMed
Summary

Direct radioimmunoassay (RIA) for human corticotrophin-releasing factor (CRF) in plasma yields inaccurate results due to interfering substances. These factors alter CRF measurements, especially in normal human plasma, unlike in equine or ovine plasma.

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Area of Science:

  • Endocrinology
  • Biochemistry
  • Immunology

Background:

  • Direct radioimmunoassay (RIA) is commonly used to measure hormone levels.
  • Human corticotrophin-releasing factor (CRF) plays a crucial role in the stress response.
  • Accurate measurement of CRF in plasma is essential for understanding various physiological and pathological conditions.

Purpose of the Study:

  • To investigate the reliability of direct RIA for measuring human CRF in peripheral plasma.
  • To identify potential interfering substances in human plasma that affect CRF measurements.
  • To compare CRF-like immunoreactivity (CRF-LI) in human, equine, and ovine plasma.

Main Methods:

  • Direct RIA using seven different antisera for human CRF.
  • Parallel dilution studies with synthetic human CRF-41.

Related Experiment Videos

  • Plasma fractionation using Sephadex G-75 and Sephacryl S-200 chromatography.
  • Methanol treatment of plasma samples.
  • Incubation of 125I-labelled human CRF-41 with human plasma.

    • Main Results:

    • Normal human plasma exhibited high CRF-like immunoreactivity (CRF-LI) that did not parallel synthetic CRF-41.
    • The majority of CRF-LI in human plasma was of large molecular size (approx. 29,000 Da).
    • Methanol treatment reduced CRF-LI in normal plasma but not in late pregnancy plasma.
    • Added labelled human CRF-41 showed altered molecular size and reduced binding in human plasma.

    Conclusions:

    • Direct RIA of unextracted human plasma for CRF provides spurious results.
    • Human plasma contains interfering factors or binding substances affecting CRF measurement.
    • These interfering substances are not consistently observed in equine or ovine plasma.
    • Further purification or modified assay methods are necessary for accurate human plasma CRF quantification.