Screening for antibodies against MICA by luminex flow cytometry

Yizhou Zou, Peter Stastny

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

Abstract

Antibodies against MICA have been found in organ transplant recipients and were found to be associated with decreased survival of kidney allografts. The MICA antibody screening assay is a Luminex-based solid phase immunoassay designed to detect IgG antibodies binding to beads pre-coated with recombinant preparations of MICA alleles. These beads coated with soluble MICA recombinant proteins including 11 common alleles have been produced in our laboratory and similar preparations have been available from commercial sources. Here, we describe the procedure of MICA antibody screening with a prepared kit, in which all the reagents were optimized and standardized. We also review how to document the quality of single MICA antigen beads using MICA-specific monoclonal antibodies, as well as quality control of the procedure and data analysis.

Original languageEnglish (US)
Title of host publicationImmunogenetics
Subtitle of host publicationMethods and Applications in Clinical Practice
EditorsFrank T. Christiansen, Brian D Tait
Pages279-288
Number of pages10
DOIs
StatePublished - Dec 1 2012

Publication series

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

Keywords

  • Antibodies
  • Luminex
  • MICA epitopes
  • Positive and negative control sera
  • Single antigen MICA beads

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Zou, Y., & Stastny, P. (2012). Screening for antibodies against MICA by luminex flow cytometry. In F. T. Christiansen, & B. D. Tait (Eds.), Immunogenetics: Methods and Applications in Clinical Practice (pp. 279-288). (Methods in Molecular Biology; Vol. 882). https://doi.org/10.1007/978-1-61779-842-9_16