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Related Experiment Videos

Rapid, homogeneous phase, liposome-based assays for total complement activity.

G Akots, J C Braman, R J Broeze

    Complement (Basel, Switzerland)
    |January 1, 1984
    PubMed
    Summary
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    A new, rapid assay quantifies complement activity using enzyme-linked liposomes. This method accurately measures complement fixation by antibodies and soluble complexes, aiding immunological research.

    Area of Science:

    • Immunology
    • Biochemistry

    Background:

    • Complement activity is crucial in immune responses.
    • Existing assays for complement activity can be complex and time-consuming.

    Purpose of the Study:

    • To develop a simple, rapid, and non-isotopic assay for determining total complement activity.
    • To adapt the assay for measuring complement-fixing antibody titers and detecting soluble antigen-antibody complexes.

    Main Methods:

    • A homogeneous, spectrophotometric assay was developed using Dnp-tagged liposomes with encapsulated enzyme.
    • Complement activity was measured by the degree of enzyme unmasking mediated by antibody and complement.
    • The assay was modified to assess complement fixation by monoclonal anti-Dnp antibodies (IgG1 and IgM) and soluble antigen-antibody complexes.

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    Main Results:

    • Complement activity in guinea pig serum was found to be proportional to concentration.
    • An IgM anti-Dnp antibody was 450-fold more effective than an IgG1 anti-Dnp antibody in mediating complement-dependent liposome damage.
    • The modified assay could detect as little as 2 pmol of Dnp antigen in soluble complexes.

    Conclusions:

    • The developed assay provides a simple, rapid, and sensitive method for quantifying total complement activity.
    • This assay is versatile and can be used to determine complement-fixing antibody titers and detect immune complexes.
    • The assay's sensitivity and ease of use make it valuable for immunological research and diagnostics.