Protein Microarray for Disease Analysis pp 81-104

Part of the Methods in Molecular Biology book series (MIMB, volume 723)

Recombinant Antigen Microarrays for Serum/Plasma Antibody Detection



Recombinant antigen arrays represent a new frontier in parallel analysis of multiple immune response profiles requiring only minute blood samples. In this article, we review the benefits and pitfalls of recombinant antigen microarrays developed for multiplexed antibody quantification. In particular, we describe the development of antigen arrays presenting a set of Y chromosome-encoded antigens, called H-Y antigens. These H-Y antigens are immunologically recognized as minor histocompatibility antigens (mHA) following allogeneic blood and organ transplantation. Clinically relevant B-cell responses against H-Y antigens have been demonstrated in male patients receiving female hematopoietic stem cell grafts and are associated with chronic graft versus host development. This chapter discusses our recombinant antigen microarray methods to measure these clinically relevant allo-antibodies.

Key words

H-Y proteins Antibodies Plasma Recombinant antigen microarrays Minor histocompatibility antigens 


  1. 1.
    Spence RP, Wright V, Ala-Aldeen DA, Turner DP, Wooldridge KG, James R (2008) Validation of virulence and epidemiology DNA microarray for identification and characterization of Staphylococcus aureus isolates. J Clin Microbiol 46:1620–1627PubMedCrossRefGoogle Scholar
  2. 2.
    Jin D, Qi H, Chen S, Zeng T, Liu Q, Wang S (2008) Simultaneous detection of six human diarrheal pathogens by using DNA microarray combined with tyramide signal amplification. J Microbiol Methods 75:365–368PubMedCrossRefGoogle Scholar
  3. 3.
    Jandu N, Ho NK, Donato KA, Karmali MA, Mascarenhas M, Duffy SP, Tailor C, Sherman PM (2009) Enterohemorrhagic Escherichia coli O157:H7 gene expression profiling in response to growth in the presence of host epithelia. PLoS One 4:e4889PubMedCrossRefGoogle Scholar
  4. 4.
    Mengual L, Burset M, Ars E, Lozano JJ, Villavicencio H, Ribal MJ, Alcaraz A (2009) DNA microarray expression profiling of bladder cancer allows identification of noninvasive diagnostic markers. J Urol 182:741–748PubMedCrossRefGoogle Scholar
  5. 5.
    Li L, Wadia P, Chen R, Kambham N, Naesens M, Sigdel TK, Miklos DB, Sarwal MM, Butte AJ (2009) Identifying compartment-specific non-HLA targets after renal transplantation by integrating transcriptome and “antibodyome” measures. Proc Natl Acad Sci U S A 106:4148–4153PubMedCrossRefGoogle Scholar
  6. 6.
    Sakuishi K, Oki S, Araki M, Porcelli SA, Miyake S, Yamamura T (2007) Invariant NKT cells biased for IL-5 production act as crucial regulators of inflammation. J Immunol 179:3452–3462PubMedGoogle Scholar
  7. 7.
    Imamichi T, Yang J, Huang DW, Brann TW, Fullmer BA, Adelsberger JW, Lempicki RA, Baseler MW, Lane HC (2008) IL-27, a novel anti-HIV cytokine, activates multiple interferon-inducible genes in macrophages. Aids 22:39–45PubMedCrossRefGoogle Scholar
  8. 8.
    Hwang SS, Kim YU, Lee W, Lee GR (2009) Differential expression of nuclear receptors in T helper cells. J Microbiol Biotechnol 19:208–214PubMedCrossRefGoogle Scholar
  9. 9.
    De Vos J, Hose D, Reme T, Tarte K, Moreaux J, Mahtouk K, Jourdan M, Goldschmidt H, Rossi JF, Cremer FW, Klein B (2006) Microarray-based understanding of normal and malignant plasma cells. Immunol Rev 210:86–104PubMedCrossRefGoogle Scholar
  10. 10.
    Yamamoto R, Sakamoto T, Nishi S, Sakai M, Morinaga T, Tamaoki T (1990) Expression of human alpha-fetoprotein in yeast. Life Sci 46:1679–1686PubMedCrossRefGoogle Scholar
  11. 11.
    Carruthers AM, Warner AJ, Michel AD, Feniuk W, Humphrey PP (1999) Activation of adenylate cyclase by human recombinant sst5 receptors expressed in CHO-K1 cells and involvement of Galphas proteins. Br J Pharmacol 126:1221–1229PubMedCrossRefGoogle Scholar
  12. 12.
    Predki PF, Mattoon D, Bangham R, Schweitzer B, Michaud G (2005) Protein microarrays: a new tool for profiling antibody cross-reactivity. Hum Antibodies 14:7–15PubMedGoogle Scholar
  13. 13.
    Mattoon D, Michaud G, Merkel J, Schweitzer B (2005) Biomarker discovery using protein microarray technology platforms: antibody-antigen complex profiling. Expert Rev Proteomics 2:879–889PubMedCrossRefGoogle Scholar
  14. 14.
    Wadia PP, Coram M, Armstrong RJ, Mindrinos M, Butte AJ, Miklos DB (2010) Antibodies specifically target AML antigen NuSAP1 after allogeneic bone marrow transplantation. Blood 115(10):2077–2087PubMedCrossRefGoogle Scholar
  15. 15.
    Marina O, Biernacki MA, Brusic V, Wu CJ (2008) A concentration-dependent analysis method for high density protein microarrays. J Proteome Res 7:2059–2068PubMedCrossRefGoogle Scholar
  16. 16.
    Stillman BA, Tonkinson JL (2000) FAST slides: a novel surface for microarrays. Biotechniques 29:630–635PubMedGoogle Scholar
  17. 17.
    MacBeath G, Schreiber SL (2000) Printing proteins as microarrays for high-throughput function determination. Science 289:1760–1763PubMedGoogle Scholar
  18. 18.
    Angenendt P, Glokler J, Murphy D, Lehrach H, Cahill DJ (2002) Toward optimized antibody microarrays: a comparison of current microarray support materials. Anal Biochem 309:253–260PubMedCrossRefGoogle Scholar
  19. 19.
    Kusnezov W, Pulli T, Witt O, Hoheisel JO (2005) Solid Supports for protien microarrays and related devices. Jones and Bartlett, Sudbury, pp 247–283Google Scholar
  20. 20.
    Paborsky LR, Dunn KE, Gibbs CS, Dougherty JP (1996) A nickel chelate microtiter plate assay for six histidine-containing proteins. Anal Biochem 234:60–65PubMedCrossRefGoogle Scholar
  21. 21.
    Zhu H, Bilgin M, Bangham R, Hall D, Casamayor A, Bertone P, Lan N, Jansen R, Bidlingmaier S, Houfek T, Mitchell T, Miller P, Dean RA, Gerstein M, Snyder M (2001) Global analysis of protein activities using proteome chips. Science 293:2101–2105PubMedCrossRefGoogle Scholar
  22. 22.
    Goulmy E (1996) Human minor histocompatibility antigens. Curr Opin Immunol 8:75–81PubMedCrossRefGoogle Scholar
  23. 23.
    Miklos DB, Kim HT, Zorn E, Hochberg EP, Guo L, Mattes-Ritz A, Viatte S, Soiffer RJ, Antin JH, Ritz J (2004) Antibody response to DBY minor histocompatibility antigen is induced after allogeneic stem cell transplantation and in healthy female donors. Blood 103:353–359PubMedCrossRefGoogle Scholar
  24. 24.
    Miklos DB, Kim HT, Miller KH, Guo L, Zorn E, Lee SJ, Hochberg EP, Wu CJ, Alyea EP, Cutler C, Ho V, Soiffer RJ, Antin JH, Ritz J (2005) Antibody responses to H-Y minor histocompatibility antigens correlate with chronic graft-versus-host disease and disease remission. Blood 105:2973–2978PubMedCrossRefGoogle Scholar
  25. 25.
    Zorn E, Miklos DB, Floyd BH, Mattes-Ritz A, Guo L, Soiffer RJ, Antin JH, Ritz J (2004) Minor histocompatibility antigen DBY elicits a coordinated B and T cell response after allogeneic stem cell transplantation. J Exp Med 199:1133–1142PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Persis P. Wadia
    • 1
  • Bita Sahaf
    • 1
  • David B. Miklos
    • 1
  1. 1.Department of Medicine, Blood and Marrow Transplantation DivisionStanford UniversityStanfordUSA

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