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Optimization and qualification of an Fc Array assay for assessments of antibodies against HIV-1/SIV.

ABSTRACT: The Fc Array is a multiplexed assay that assesses the Fc domain characteristics of antigen-specific antibodies with the potential to evaluate up to 500 antigen specificities simultaneously. Antigen-specific antibodies are captured on antigen-conjugated beads and their functional capacity is probed via an array of Fc-binding proteins including antibody subclassing reagents, Fc? receptors, complement proteins, and lectins. Here we present the results of the optimization and formal qualification of the Fc Array, performed in compliance with Good Clinical Laboratory Practice (GCLP) guidelines. Assay conditions were optimized for performance and reproducibility, and the final version of the assay was then evaluated for specificity, accuracy, precision, limits of detection and quantitation, linearity, range and robustness.

SUBMITTER: Brown EP

PROVIDER: S-EPMC5851662 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature

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Optimization and qualification of an Fc Array assay for assessments of antibodies against HIV-1/SIV.

Brown Eric P EP Weiner Joshua A JA Lin Shu S Natarajan Harini H Normandin Erica E Barouch Dan H DH Alter Galit G Sarzotti-Kelsoe Marcella M Ackerman Margaret E ME

Journal of immunological methods 20180131

The Fc Array is a multiplexed assay that assesses the Fc domain characteristics of antigen-specific antibodies with the potential to evaluate up to 500 antigen specificities simultaneously. Antigen-specific antibodies are captured on antigen-conjugated beads and their functional capacity is probed via an array of Fc-binding proteins including antibody subclassing reagents, Fcγ receptors, complement proteins, and lectins. Here we present the results of the optimization and formal qualification of ...[more]

PMID: 29395167

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