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A solid-phase ELISA for human galactosyltransferase.

B Verdon, T Mandel, E G Berger

    Journal of Immunological Methods
    |November 26, 1982
    PubMed
    Summary
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    Researchers developed a new enzyme-linked immunosorbent assay (ELISA) to measure human milk galactosyltransferase. This assay accurately quantifies the enzyme in milk and other bodily fluids.

    Area of Science:

    • Biochemistry
    • Immunology
    • Enzymology

    Background:

    • Human milk galactosyltransferase is a key enzyme in lactose synthesis.
    • Accurate quantification of this enzyme is important for understanding its role in infant nutrition and development.
    • Existing methods for enzyme quantification may lack sensitivity or specificity.

    Purpose of the Study:

    • To develop and validate a sensitive solid-phase enzyme-linked immunosorbent assay (ELISA) for human milk galactosyltransferase.
    • To determine the detection limit and precision of the developed ELISA.
    • To quantify galactosyltransferase levels in human milk samples.

    Main Methods:

    • Development of a solid-phase ELISA using monospecific rabbit anti-human milk galactosyltransferase antibodies.

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  • Assay design based on competition with a galactosyltransferase-phosphatase conjugate.
  • Quantification of enzyme in dialyzed milk samples.

    • Main Results:

    • The ELISA demonstrated a detection limit of 30 ng/ml.
    • Intra-assay and inter-assay coefficients of variation were less than 7.7% and 6.8-18.8%, respectively.
    • The average galactosyltransferase content in human milk was determined to be 61 ± 19 μg/ml.

    Conclusions:

    • The developed ELISA is a reliable and sensitive method for quantifying human milk galactosyltransferase.
    • The assay is suitable for analyzing enzyme levels in both milk and partially purified enzyme preparations from body fluids.
    • This assay facilitates further research into the physiological roles of galactosyltransferase.