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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bahram S, Bresnahan M, Geraghty DE et al (1994) A second lineage of mammalian major histocompatibility complex class I genes. Proc Natl Acad Sci 91:6259–6263
Zou Y, Heinemann FM, Grosse-Wilde H et al (2006) Detection of anti-MICA antibodies in patients awaiting kidney transplantation, during the post-transplant course, and in eluates from rejected kidney allografts by Luminex flow cytometry. Hum Immunol 67:230–237
Terasaki PI, Ozawa M, Castro R (2007) Four-year follow-up of a prospective trial of HLA and MICA antibodies on kidney graft survival. Am J Transplant 7:408–415
Zou Y, Stastny P, Süsal C et al (2007) Antibodies against MICA antigens and kidney-transplant rejection. N Engl J Med 357:1293–1300
Zou Y, Qin Z, Silveus A et al (2009) Polymorphisms of MICA recognized by human alloantibodies. Immunogenetics 61:91–100
Duquesnoy RJ, Mostecki J, Hariharan J et al (2008) Structurally based epitope analysis of major histocompatibility complex class I-related chain A (MICA) antibody specificity patterns. Hum Immunol 69:826–832
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Zou, Y., Stastny, P. (2012). Screening for Antibodies Against MICA by Luminex Flow Cytometry. In: Christiansen, F., Tait, B. (eds) Immunogenetics. Methods in Molecular Biology, vol 882. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-842-9_16
Download citation
DOI: https://doi.org/10.1007/978-1-61779-842-9_16
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-841-2
Online ISBN: 978-1-61779-842-9
eBook Packages: Springer Protocols