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ID: 174921.0, MPI für molekulare Genetik / Department of Vertebrate Genomics
A nonredundant human protein chip for antibody screening and serum profiling
Authors:Lueking, Angelika; Possling, Alexandra; Huber, Otmar; Beveridge, Allan; Horn, Martin; Eickhoff, Holger; Schuchardt, Johannes; Lehrach, Hans; Cahill, Dolores J.
Date of Publication (YYYY-MM-DD):2003-12
Title of Journal:Molecular & Cellular Proteomics
Journal Abbrev.:Mol. Cell. Proteomics
Issue / Number:12
Start Page:1342
End Page:1349
Copyright:© 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
Review Status:not specified
Audience:Experts Only
Abstract / Description:There is burgeoning interest in protein microarrays, but a source of thousands of nonredundant, purified proteins was not previously available. Here we show a glass chip containing 2413 nonredundant purified human fusion proteins on a polymer surface, where densities up to 1600 proteins/cm2 on a microscope slide can be realized. In addition, the polymer coating of the glass slide enables screening of protein interactions under nondenaturing conditions. Such screenings require only 200-µl sample volumes, illustrating their potential for high-throughput applications. Here we demonstrate two applications: the characterization of antibody binding, specificity, and cross-reactivity; and profiling the antibody repertoire in body fluids, such as serum from patients with autoimmune diseases. For the first application, we have incubated these protein chips with anti-RGSHis6, anti-GAPDH, and anti-HSP90ß antibodies. In an initial proof of principle study for the second application, we have screened serum from alopecia and arthritis patients. With analysis of large sample numbers, identification of disease-associated proteins to generate novel diagnostic markers may be possible.
Comment of the Author/Creator:Originally published In Press as doi:10.1074/mcp.T300001-MCP200 on September 29, 2003.
External Publication Status:published
Document Type:Article
Version Comment:Automatic journal name synchronization
Communicated by:Hans Lehrach
Affiliations:MPI für molekulare Genetik
External Affiliations:Royal Coll Surgeons Ireland, Ctr Human Proteom, Dublin 2, Ireland.; Protagen AG, D-44227 Dortmund, Germany.; Max Planck Inst Mol Genet, D-14195 Berlin, Germany.; Free Univ Berlin, Klinikum Benjamin Franklin, Inst Clin Chem & Pathobiochem, D-12200 Berlin, Germany.; Scien AG, D-12489 Berlin, Germany.; MicroDiscovery GmbH, D-10405 Berlin, Germany.
Identifiers:ISI:000187250900009 [ID No:1]
ISSN:1535-9476 [ID No:2]
DOI:10.1074/mcp.T300001-MCP200 [ID No:3]
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