Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 473, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.jim.2019.07.002
Keywords
ADCD; Complement; Antibody-dependent effector function; High-throughput; Fc receptor
Categories
Funding
- NIH, United States [AI080289, AI102660-01, AI129797-01]
- Bill and Melinda Gates Foundation, United States [OPP1114729, OPP1146996, OPP1032817]
- Massachusetts General Hospital ECOR, United States
- Harvard University Center for AIDS Research, United States [P30 AI060354-02]
- Ragon Institute of MGH, MIT, and Harvard, United States
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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.
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