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

• Published in: Journal of immunological methods Vol. 473; p. 112630
• Main Authors: Fischinger, Stephanie, Fallon, Jonathan K., Michell, Ashlin R., Broge, Thomas, Suscovich, Todd J., Streeck, Hendrik, Alter, Galit
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.10.2019; Elsevier
• Subjects: ADCD; Animals; antibodies; Antibodies - immunology; Antibody-dependent effector function; antigen-antibody complex; antigens; Complement; Complement Activation; Complement System Proteins - physiology; Fc receptor; Flow Cytometry; Guinea Pigs; High-throughput; High-Throughput Screening Assays - methods; Humans; innate immunity; pathogens; Reproducibility of Results; High-throughput; ADCD; Complement; Antibody-dependent effector function; Fc receptor
• ISSN: 0022-1759; 1872-7905; 1872-7905
• DOI: 10.1016/j.jim.2019.07.002

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. •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