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


Journal

Journal of immunological methods
ISSN: 1872-7905
Titre abrégé: J Immunol Methods
Pays: Netherlands
ID NLM: 1305440

Informations de publication

Date de publication:
10 2019
Historique:
received: 29 12 2018
revised: 21 05 2019
accepted: 08 07 2019
pubmed: 14 7 2019
medline: 7 5 2020
entrez: 14 7 2019
Statut: ppublish

Résumé

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.

Identifiants

pubmed: 31301278
pii: S0022-1759(18)30463-0
doi: 10.1016/j.jim.2019.07.002
pmc: PMC6722412
pii:
doi:

Substances chimiques

Antibodies 0
Complement System Proteins 9007-36-7

Types de publication

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

Langues

eng

Sous-ensembles de citation

IM

Pagination

112630

Subventions

Organisme : NIAID NIH HHS
ID : U19 AI135995
Pays : United States
Organisme : NIAID NIH HHS
ID : P30 AI060354
Pays : United States

Informations de copyright

Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.

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Auteurs

Stephanie Fischinger (S)

Ragon Institute of MGH, Harvard and MIT, Cambridge 02139, USA; University of Duisburg-Essen, Essen 47057, Germany.

Jonathan K Fallon (JK)

Ragon Institute of MGH, Harvard and MIT, Cambridge 02139, USA.

Ashlin R Michell (AR)

Ragon Institute of MGH, Harvard and MIT, Cambridge 02139, USA.

Thomas Broge (T)

Ragon Institute of MGH, Harvard and MIT, Cambridge 02139, USA.

Todd J Suscovich (TJ)

Ragon Institute of MGH, Harvard and MIT, Cambridge 02139, USA.

Hendrik Streeck (H)

University of Duisburg-Essen, Essen 47057, Germany.

Galit Alter (G)

Ragon Institute of MGH, Harvard and MIT, Cambridge 02139, USA. Electronic address: galter@mgh.harvard.edu.

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Classifications MeSH