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Immunogenetics pp 279-288 | Cite as

Screening for Antibodies Against MICA by Luminex Flow Cytometry

  • Yizhou Zou
  • Peter Stastny
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 882)

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.

Key words

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

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Division of Transplant Immunology, Department of Internal MedicineUT Southwestern Medical CentreDallasUSA

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