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Stability studies on human pituitary prolactin.

F Nyberg, P Bergman, L Wide

    Upsala Journal of Medical Sciences
    |January 1, 1985
    PubMed
    Summary
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    Human pituitary prolactin stability was assessed. Alkaline conditions caused deamidation, reducing immunoactivity, while serum accelerated this change, suggesting enzymatic activity. Ethylene glycol ensured long-term stability.

    Area of Science:

    • Biochemistry
    • Endocrinology

    Background:

    • Human pituitary prolactin exists as differently charged isomers.
    • Understanding prolactin stability is crucial for its accurate measurement and therapeutic use.

    Purpose of the Study:

    • To investigate the stability of human pituitary prolactin isomers under various conditions.
    • To elucidate the mechanisms of prolactin alteration and identify factors affecting its stability and immunoactivity.

    Main Methods:

    • Analytical electrophoresis and radioimmunoassay were employed to monitor prolactin changes.
    • Quantitative kinetic studies were performed to determine the rate constants of observed alterations.
    • Prolactin stability was assessed under different pH, ionic strength, and storage conditions, including the presence of ethylene glycol and human serum.

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    Main Results:

    • At alkaline pH (10), prolactin isomers underwent conversion to faster-migrating forms, likely due to deamidation, which correlated with decreased immunoactivity.
    • The rate of this deamidation decreased with increasing acidity of prolactin components.
    • In human serum at neutral pH, prolactin conversion occurred at a rate approximately four times higher than in saline, suggesting enzymatic involvement.
    • Prolactin stability was concentration-dependent and influenced by buffer ionic strength.
    • At -20°C with 50% ethylene glycol and protein concentrations of 0.1-2 mg/ml, prolactin remained stable for years, both immunologically and electrophoretically.

    Conclusions:

    • The observed alteration at pH 10 is likely due to non-enzymic deamidation.
    • Enzymatic conversion of prolactin occurs in human serum.
    • Optimal storage conditions, including the use of ethylene glycol at low temperatures and specific protein concentrations, are essential for preserving prolactin integrity over extended periods.