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Related Experiment Videos

A novel assay for serum complement activity: C42 generation assay.

E Kobayashi1, E Kitano, H Kitamura

  • 1Department of Medical Technology, Osaka Prefectural College of Health Sciences, Osaka, Japan.

International Archives of Allergy and Immunology
|October 26, 1999
PubMed
Summary

A new C42 generation assay offers a rapid and economical method to analyze serum complement activity. This functional assay provides valuable insights into low CH50 levels, complementing traditional tests.

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Area of Science:

  • Immunology
  • Clinical Chemistry
  • Complement System Analysis

Background:

  • Current clinical laboratory tests for serum complement, such as C3, C4, and CH50, are often insufficient for analyzing low CH50 serum.
  • Limitations exist in the diagnostic capacity of standard immunochemical and functional complement assays.

Purpose of the Study:

  • To introduce and evaluate a novel C42 generation assay for assessing serum complement activity.
  • To provide a more comprehensive analysis of serum complement function, particularly for cases with decreased CH50.

Main Methods:

  • Development of a new assay based on the hemolysis of sensitized sheep erythrocytes (EA).
  • The assay involves two stages: generation of C42 (classical pathway C3 convertase) and subsequent hemolysis by C3-C9.

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  • Utilizes two sera sequentially, with the second serum providing C3-C9 in the presence of EDTA.

    • Main Results:

    • The C42 generation assay allows for the evaluation of two distinct serum complement activities: C1, C4, and C2 combined, and C3-C9 combined.
    • Hemolysis levels directly correlate with the activity of these complement components.
    • The assay proved helpful in analyzing clinical cases with decreased CH50 serum, with several examples presented.

    Conclusions:

    • The C42 generation assay is a valuable functional assay for serum complement.
    • It provides additional information beyond the standard CH50 assay.
    • The assay is rapid, economical, and does not require intermediate cells or isolated complement components, making it suitable for clinical settings.