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

J Immunol Methods. 2018 Apr:455:24-33. doi: 10.1016/j.jim.2018.01.013. Epub 2018 Jan 31.

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, Fcγ 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.

Publication types

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

MeSH terms

  • Animals
  • Antigens, Viral / immunology
  • Antigens, Viral / metabolism
  • Complement System Proteins / immunology
  • Complement System Proteins / metabolism
  • HIV Infections / diagnosis*
  • HIV Infections / immunology
  • HIV-1 / physiology*
  • Humans
  • Immunoassay / methods*
  • Immunoglobulin Fc Fragments / analysis*
  • Lectins / immunology
  • Lectins / metabolism
  • Macaca mulatta
  • Microspheres
  • Practice Guidelines as Topic
  • Protein Binding
  • Reference Standards
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Simian Acquired Immunodeficiency Syndrome / diagnostic imaging*
  • Simian Acquired Immunodeficiency Syndrome / immunology
  • Simian Immunodeficiency Virus / physiology*

Substances

  • Antigens, Viral
  • Immunoglobulin Fc Fragments
  • Lectins
  • Complement System Proteins