Practical enzyme immunoassay for prolactin in human serum.

Area of Science:

• Biochemistry
• Immunology
• Clinical Chemistry

Background:

• Prolactin is a crucial hormone regulating numerous physiological processes.
• Accurate measurement of serum prolactin is essential for diagnosing and monitoring various endocrine disorders.
• Existing methods like radioimmunoassay have limitations, necessitating the development of alternative assays.

Purpose of the Study:

• To develop and validate a novel competitive enzyme immunoassay (EIA) for the quantitative determination of prolactin in human serum.
• To assess the sensitivity, accuracy, and correlation of the developed EIA with established radioimmunoassay (RIA) methods.

Main Methods:

• A competitive enzyme immunoassay was designed using anti-prolactin antibodies and beta-D-galactosidase-labeled prolactin.
• Serum samples were incubated with antibodies and labeled prolactin, followed by separation of bound and unbound fractions using a thiol-disulfide interchange reaction.
• Enzyme activity measurement was employed to quantify the amount of bound prolactin, thereby determining serum prolactin concentration.

Main Results:

• The developed enzyme immunoassay demonstrated a sensitivity of 2.5 ng/ml.
• The assay exhibited comparable sensitivity to traditional radioimmunoassay.
• A strong correlation was observed between the results obtained from the enzyme immunoassay and radioimmunoassay (r = 0.98, slope = 1.26, y-intercept = -12.8 ng/ml).

Conclusions:

• The developed competitive enzyme immunoassay provides a sensitive and accurate method for measuring serum prolactin.
• This EIA serves as a viable alternative to radioimmunoassay for clinical diagnostics.
• The assay's high correlation with RIA validates its reliability in clinical settings.