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

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

Author information

1
Ragon Institute of MGH, Harvard and MIT, Cambridge 02139, USA; University of Duisburg-Essen, Essen 47057, Germany.
2
Ragon Institute of MGH, Harvard and MIT, Cambridge 02139, USA.
3
University of Duisburg-Essen, Essen 47057, Germany.
4
Ragon Institute of MGH, Harvard and MIT, Cambridge 02139, USA. Electronic address: galter@mgh.harvard.edu.

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

PMID:
31301278
DOI:
10.1016/j.jim.2019.07.002
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