Advertisement

Protein Arrays II: Antigen Arrays

  • Yulin Yuan
  • Hongting Wang
  • Zuan-Tao Lin
  • Xia Hong
  • Mikala Heon
  • Tianfu WuEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1654)

Abstract

Antigen arrays are fabricated using various antigens such as DNA, histones, synthetic peptides, recombinant proteins, or cell extracts to detect autoantibodies in autoimmune diseases, alloantibodies in transplantation, drug-induced antibodies or cancer-induced antibodies in blood or cell culture supernatant. In this protocol, we will provide a step-by-step executable procedure to perform antigen arrays, including antigen preparation and printing, blocking, sample loading, array detection, imaging, and data analysis.

Key words

Antigen arrays Autoantibody profiling Biomarker 

Notes

Acknowledgments

This work was partly supported by a grant from the Lupus Research Institute to T.W. and a startup fund from the University of Houston to T.W.

References

  1. 1.
    Prechl J, Papp K, Erdei A (2010) Antigen microarrays: descriptive chemistry or functional immunomics? Trends Immunol 31(4):133–137CrossRefPubMedGoogle Scholar
  2. 2.
    Li QZ, Xie C, Wu T, Mackay M, Aranow C, Putterman C, Mohan C (2005) Identification of autoantibody clusters that best predict lupus disease activity using glomerular proteome arrays. J Clin Invest 115(12):3428–3439. doi: 10.1172/JCI23587 CrossRefPubMedGoogle Scholar
  3. 3.
    Ramachandran N, Raphael JV, Hainsworth E, Demirkan G, Fuentes MG, Rolfs A, Hu Y, LaBaer J (2008) Next-generation high-density self-assembling functional protein arrays. Nat Methods 5(6):535–538CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Tabakman SM, Lau L, Robinson JT, Price J, Sherlock SP, Wang H, Zhang B, Chen Z, Tangsombatvisit S, Jarrell JA, Utz PJ, Dai H (2011) Plasmonic substrates for multiplexed protein microarrays with femtomolar sensitivity and broad dynamic range. Nat Commun 2:466. doi: 10.1038/ncomms1477 CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Zhang B, Kumar RB, Dai H, Feldman BJ (2014) A plasmonic chip for biomarker discovery and diagnosis of type 1 diabetes. Nat Med 20(8):948–953. doi: 10.1038/nm.3619 CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Papp K, Prechl J (2012) The use of antigen microarrays in antibody profiling. Methods Mol Biol 815:175–185CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Yulin Yuan
    • 1
    • 2
  • Hongting Wang
    • 1
    • 3
  • Zuan-Tao Lin
    • 1
  • Xia Hong
    • 1
    • 4
  • Mikala Heon
    • 1
  • Tianfu Wu
    • 1
    Email author
  1. 1.Department of Biomedical EngineeringUniversity of HoustonHoustonUSA
  2. 2.Department of Clinical LaboratoryThe People’s Hospital of Guangxi Zhuang Autonomous RegionNanningChina
  3. 3.National Pharmacology Laboratory of Chinese MedicineCollege of Basic Medical Sciences, Wannan Medical CollegeWuhuChina
  4. 4.Department of NursingFujian Health CollegeFuzhouChina

Personalised recommendations