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

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

Author information

1
Thayer School of Engineering, Dartmouth College, Hanover, NH 03755, United States.
2
Ragon Institute of MGH, MIT, Harvard University, 149 13th St, Charlestown, MA 02129, United States; Beth Israel Deaconess Medical Center, Boston, MA 02215, United States.
3
Ragon Institute of MGH, MIT, Harvard University, 149 13th St, Charlestown, MA 02129, United States.
4
Duke University Medical Center, 2812 Erwin Rd., Suite 301, Erwin Terrace II, Durham, NC 27705, United States.
5
Thayer School of Engineering, Dartmouth College, Hanover, NH 03755, United States. Electronic address: margaret.e.ackerman@dartmouth.edu.

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.

PMID:
29395167
PMCID:
PMC5851662
DOI:
10.1016/j.jim.2018.01.013
[Indexed for MEDLINE]
Free PMC Article

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