The complement-mediated prozone effect in the Luminex single-antigen bead assay and its impact on HLA antibody determination in patient sera

Int J Immunogenet. 2013 Jun;40(3):171-7. doi: 10.1111/j.1744-313X.2012.01147.x. Epub 2012 Aug 23.

Abstract

The Luminex xMAP system has become an important tool for HLA antibody screening and identification in sera of transplant patients. Recently, the Luminex single antigen bead assay was shown to be prone to an artefact, the so called prozone phenomenon: Sera with high titer HLA antibodies gave negative results when tested neat, but reacted strongly positive after 1:10 dilution. We also observed such a phenomenon and found that it was most likely caused by the complement component 1 (C1) by competitively displacing the detection antibodies. In this article we review the complement-mediated prozone effect and other mechanisms of interference with solid phase assays, and we discuss possible consequences for HLA antibody testing with the Luminex SAB assay.

Publication types

  • Review

MeSH terms

  • Antibodies / blood*
  • Antibodies / immunology*
  • Antigens / immunology
  • Complement C1 / immunology
  • Complement C4 / immunology
  • Complement System Proteins / immunology*
  • HLA Antigens / immunology*
  • Histocompatibility Testing / methods*
  • Histocompatibility Testing / standards*
  • Humans
  • Immunoassay / methods

Substances

  • Antibodies
  • Antigens
  • Complement C1
  • Complement C4
  • HLA Antigens
  • Complement System Proteins