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Classical pathway evaluation

P C Giclas1

  • 1National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado, USA.

Current Protocols in Immunology
|April 25, 2008
PubMed

Insights

This study details complement system assays for assessing classical pathway function and protein levels. Methods include CH(50) assays and radial immunodiffusion for comprehensive complement analysis.

Area of Science:

  • Immunology
  • Biochemistry

Background:

  • The classical complement pathway is crucial for immune responses.
  • Accurate assessment of complement function and protein levels is vital for diagnosing immunodeficiencies and autoimmune diseases.

Purpose of the Study:

  • To describe standardized assay methods for evaluating the classical complement pathway's functional status.
  • To provide protocols for quantitating individual complement component proteins.

Main Methods:

  • Two CH(50) assay variations (tube-based and microtiter plate) for total classical pathway function.
  • Radial immunodiffusion (RID) methods for measuring circulating complement protein concentrations.
  • Support protocols for preparing reagents like erythrocyte-antibody complexes (EA) and titrating hemolysin.

Main Results:

  • Established procedures for functional assessment of the entire classical complement cascade (C1qrs to C9).
  • Quantification of key complement proteins using RID, offering an alternative to functional assays.
  • Method for determining C1-esterase inhibitor functional status and antibody concentrations for RID.

Conclusions:

  • The described methods provide a robust framework for comprehensive analysis of the classical complement pathway.
  • These assays are essential tools for immunological research and clinical diagnostics.
  • Standardized protocols ensure reliable and reproducible complement system evaluation.

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