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The quantitation of alternative pathway complement function by timed lysis assay.

B M Jones

    Journal of Immunological Methods
    |January 1, 1979
    PubMed
    Summary
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    A novel timed lysis assay accurately quantifies human serum haemolytic complement activity. This method provides reliable classical and alternative pathway complement function measurements, comparable to established techniques.

    Area of Science:

    • Immunology
    • Biochemistry

    Background:

    • Complement system plays a crucial role in innate and adaptive immunity.
    • Accurate quantification of complement activity is essential for diagnosing immune disorders.
    • Existing methods for assessing complement function can be time-consuming or complex.

    Purpose of the Study:

    • To develop and validate a simple timed lysis assay for quantifying haemolytic complement activity in human serum.
    • To assess both classical and alternative complement pathway functions using this assay.
    • To compare the results of the timed lysis assay with established complement activity measurement techniques.

    Main Methods:

    • A timed lysis assay was developed to measure the time required for 50% lysis of antibody-coated sheep erythrocytes (classical pathway) and rabbit erythrocytes (alternative pathway).

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  • Calibration curves were generated using serial dilutions of standard human serum to convert lysis times into CH50 U/ml.
  • The timed lysis assay results were compared with those obtained from traditional dilution methods (Mayer, 1971; Platts-Mills & Ishizaka, 1974).

    • Main Results:

    • The timed lysis assay demonstrated consistent quantification of haemolytic complement activity.
    • Comparable results were obtained when comparing the timed lysis assay with established classical and alternative pathway assays.
    • The assay effectively measures both classical and alternative complement pathway functions.

    Conclusions:

    • The described timed lysis assay is a simple and reliable method for quantifying haemolytic complement activity in human serum.
    • This assay provides accurate measurements of both classical and alternative complement pathway functions.
    • The timed lysis assay offers a valuable alternative to existing methods for complement activity assessment.