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A semi-automated microassay for complement activity.

C C Liu1, J D Young

  • 1Laboratory of Cellular Physiology and Immunology, Rockefeller University, New York 10021.

Journal of Immunological Methods
|November 10, 1988
PubMed
Summary
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A novel microassay simplifies measuring serum complement activity. This automated method uses light scattering for rapid, semi-quantitative screening of hemolytic activity in many samples.

Area of Science:

  • Immunology
  • Biochemistry

Background:

  • Serum complement activity is crucial for immune response.
  • Traditional methods for assessing complement hemolytic activity are often time-consuming and complex.
  • A need exists for a simpler, automated assay for complement diagnostics.

Purpose of the Study:

  • To describe a simple, automated microassay for quantifying serum complement-dependent hemolytic activity.
  • To validate the new method against the traditional CH50 titration assay.
  • To establish the utility of this assay for rapid, large-scale screening of complement function.

Main Methods:

  • The assay utilizes changes in light scattering properties of sheep erythrocytes upon hemolysis.
  • A fixed volume of serum specimen and erythrocytes are used in a single experimental step.

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  • Spectrophotometric readings are taken at 700 nm using a microplate reader.

    • Main Results:

    • The measured absorbance showed a proportional correlation with the extent of hemolysis.
    • Excellent correlation was observed between the results from the new microassay and the traditional CH50 titration method.
    • The assay is simple, automated, and suitable for high-throughput analysis.

    Conclusions:

    • This automated microassay provides a simple and efficient method for assessing serum complement hemolytic activity.
    • The assay's correlation with the CH50 method supports its diagnostic utility.
    • It is well-suited for rapid, semi-quantitative screening of complement function in numerous serum specimens.