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

https://doi.org/10.1016/j.jim.2019.07.002Get rights and content
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Highlights

  • Optimized flow-based assay for the detection of antibody-mediated complement deposition

  • Robust, rapid and reproducible high-throughput bead-based assay applicable to various diseases, including HIV and influenza

  • Lot controlled complement is a controlled source for exogenous complement that correlates with human complement activity

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
Complement
Antibody-dependent effector function
High-throughput
Fc receptor

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Manuscript has not been submitted/accepted elsewhere, all authors contributed significantly to the work, have seen/approved the manuscript