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Related Experiment Videos

Partial purification of human lymphocyte activating factor.

L B Lachman, S O Page, R S Metzgar

    Preparative Biochemistry
    |January 1, 1980
    PubMed
    Summary
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    Researchers purified Lymphocyte Activating Factor (LAF), a T lymphocyte stimulant, using isoelectric focusing. This method effectively separated LAF from human serum proteins, yielding a highly active purified sample.

    Area of Science:

    • Immunology
    • Biochemistry
    • Cell Biology

    Background:

    • Lymphocyte Activating Factor (LAF) stimulates T lymphocytes.
    • LAF is released by human monocytes stimulated with lipopolysaccharide.
    • Purification requires separating LAF from human serum proteins.

    Purpose of the Study:

    • To develop an effective purification method for Lymphocyte Activating Factor (LAF).
    • To separate low molecular weight (MW) LAF from human serum proteins.
    • To obtain a highly active purified sample of LAF.

    Main Methods:

    • Human monocytes cultured for 18-24 hours in medium with 5% human serum and lipopolysaccharide.
    • Hollow fiber ultrafiltration for initial separation of low MW LAF.
    • Isoelectric focusing (IEF) for final purification of LAF.

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    Main Results:

    • Hollow fiber ultrafiltration yielded only 20% of original LAF activity.
    • Isoelectric focusing (IEF) efficiently separated LAF from all human serum traces.
    • IEF resulted in a purified sample with no measurable protein and high biological activity.

    Conclusions:

    • Isoelectric focusing is an effective method for purifying Lymphocyte Activating Factor (LAF).
    • The IEF-purified LAF is highly active in biological assays.
    • This purification strategy yields a cleaner, more potent LAF preparation.