A high-throughput, bead-based, antigen-specific assay to assess the ability of antibodies to induce complement activation

J Immunol Methods. 2019 Oct;473:112630. doi: 10.1016/j.jim.2019.07.002. Epub 2019 Jul 10.

Abstract

The complement system plays a critical role in innate immune defense against pathogens, both via non-specific direct pathogen recognition and killing or via antigen-specific indirect recruitment by complement fixing antibodies. While various assays for measuring complement activation have been developed, few provide a high-throughput, sample-sparing approach to interrogate the qualitative differences in the ability of antibodies to drive complement activation. Here we present a high-throughput, sample-sparing, bead-based assay to evaluate antigen-specific antibody-dependent complement activation against nearly any antigen. Optimization of buffer composition, kinetics of immune complex formation, as well as complement source all contribute critically to the development of a robust, highly flexible and high-throughput approach to analyze antibody-dependent complement deposition (ADCD). Thus, the optimized bead-based, antigen-specific assay represents a simple, highly adaptable platform to profile antibody-dependent complement activation across pathogens and diseases.

Keywords: ADCD; Antibody-dependent effector function; Complement; Fc receptor; High-throughput.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / immunology*
  • Complement Activation*
  • Complement System Proteins / physiology
  • Flow Cytometry
  • Guinea Pigs
  • High-Throughput Screening Assays / methods*
  • Humans
  • Reproducibility of Results

Substances

  • Antibodies
  • Complement System Proteins