HLA antibody detection and characterization by solid phase immunoassays: Methods and pitfalls

Andrea A. Zachary, Renato M. Vega, Donna P. Lucas, Mary S. Leffell

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Solid phase immunoassays for the detection and characterization of HLA-specific antibodies provide greatly increased sensitivity, specificity, and time and reagent efficiency, compared to the traditionally used cell-based methods. Testing is performed using commercially available test kits. The assays are of two general types: enzyme-linked immunosorbent assays and multianalyte bead. The types vary in both sensitivity and equipment requirements. While these assays afford great improvement over the cell-based assays, they can be confounded by interference from substances within the serum that result in high background reactivity. The high sensitivity of the assays also makes them more susceptible to environmental factors and operator variability. The user must be aware of the capabilities of the various formats, the factors that can affect test results, and lot to lot variability of any single product. Knowledge of the characteristics of each product and thorough and accurate analysis of the results are essential to the utility of these assays.

Original languageEnglish (US)
Title of host publicationImmunogenetics
Subtitle of host publicationMethods and Applications in Clinical Practice
EditorsFrank T. Christiansen, Brian D Tait
Pages289-308
Number of pages20
DOIs
StatePublished - Dec 1 2012

Publication series

NameMethods in Molecular Biology
Volume882
ISSN (Print)1064-3745

Keywords

  • Enzyme-linked immunosorbent assays
  • HLA antibody
  • HLA phenotype panel
  • Luminex
  • Multianalyte bead assays
  • Pooled antigen panel
  • Single antigen panel
  • Solid phase immunoassays

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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