title: High-throughput screening for native autoantigen-autoantibody complexes using antibody microarrays.
abstract: We report on a novel, high-dimensional method to detect autoantibodies that are complexed with their natural autoantigens. Specifically, autoantibody-autoantigen complexes in serum or plasma are directly incubated onto a high-density antibody microarray. Detection of the bound autoantibody-antigen complex is made via fluorescently labeled antihuman immunoglobulin G or other immunoglobulin isotype secondary antibodies and quantification in a microarray scanner. Uncomplexed antibodies do not interfere with this assay. The whole process is very rapid and applicable for high-throughput screening without the need for production of proteins or immunoglobulin purification from the samples. Using these methods, we found that plasma from healthy individuals contains hundreds of autoantibodies complexed with cellular proteins. Thus, this highly sensitive, multiplex method is capable of discovering new autoantibody-antigen or circulating immune complexes, many of which will likely be useful for disease detection and characterization.
authors.affiliation: Translational Research Program, Human Biology and Public Health Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, Seattle, Washington 98109, USA.
authors.forename: Jung-hyun
authors.initials: JH
authors.lastname: Rho
authors.forename: Paul D
authors.initials: PD
authors.lastname: Lampe
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry*; D001323:Autoantibodies / Q000737:chemistry*; D001324:Autoantigens / Q000097:blood / Q000737:chemistry* / Q000276:immunology; D014408:Biomarkers, Tumor / Q000097:blood / Q000737:chemistry* / Q000276:immunology; D016022:Case-Control Studies; D003110:Colonic Neoplasms / Q000097:blood / Q000276:immunology; D006801:Humans; D007163:Immunosorbent Techniques; D040081:Protein Array Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013194:Staining and Labeling
chemical_list: D055910:Antibodies, Immobilized; D001323:Autoantibodies; D001324:Autoantigens; D014408:Biomarkers, Tumor
references.citation: Cancer Res. 2000 Apr 1;60(7):1777-88
references.pmid: 10766157
references.citation: Cancer Immunol Immunother. 2010 Sep;59(9):1389-400
references.pmid: 20514540
references.citation: Clin Chem. 2011 Jun;57(6):905-9
references.pmid: 21482748
references.citation: Curr Protoc Protein Sci. 2011 Apr;Chapter 27:Unit27.2
references.pmid: 21488044
references.citation: Clin Chem. 2010 Dec;56(12):1889-92
references.pmid: 20943848
references.citation: Nat Biotechnol. 2011 May 22;29(6):535-41
references.pmid: 21602805
references.citation: Cancer Immunol Immunother. 1988;26(2):180-4
references.pmid: 3282655
references.citation: Biomarkers. 2012 Jun;17(4):362-71
references.pmid: 22612312
references.citation: Eur J Cancer. 2010 Jan;46(1):170-9
references.pmid: 19926475
references.citation: Int J Cancer. 2000 Jun 1;86(5):702-12
references.pmid: 10797294
references.citation: Methods Mol Med. 2005;109:137-54
references.pmid: 15585919
references.citation: J Cell Biol. 2007 Dec 17;179(6):1301-9
references.pmid: 18086922
references.citation: Am J Clin Oncol. 1999 Dec;22(6):542-9
references.pmid: 10597736
references.citation: Mol Cell Proteomics. 2009 Oct;8(10):2382-95
references.pmid: 19638618
references.citation: Mol Oncol. 2007 Dec;1(3):313-20
references.pmid: 19383305
references.citation: Nat Struct Biol. 1997 May;4(5):374-81
references.pmid: 9145108
references.citation: J Cell Mol Med. 2011 Oct;15(10):2013-24
references.pmid: 21651719
references.citation: Methods Enzymol. 2011;500:151-63
references.pmid: 21943897
references.citation: Nat Protoc. 2006;1(1):452-60
references.pmid: 17406268
references.citation: Proc Natl Acad Sci U S A. 2003 Oct 14;100(21):12361-6
references.pmid: 14519847
references.citation: Cancer Lett. 2009 Aug 28;281(2):144-50
references.pmid: 19304375
references.citation: J Proteome Res. 2005 Jul-Aug;4(4):1123-33
references.pmid: 16083262
references.citation: Cancer Detect Prev. 2007;31(5):402-7
references.pmid: 18035503
references.citation: J Proteome Res. 2011 Jan 7;10(1):85-96
references.pmid: 20977275
references.citation: Mol Cell Proteomics. 2010 Jul;9(7):1449-60
references.pmid: 20467042
references.citation: Cancer Lett. 2010 Mar 1;289(1):32-9
references.pmid: 19683863