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  A nonredundant human protein chip for antibody screening and serum profiling

Lueking, A., Possling, A., Huber, O., Beveridge, A., Horn, M., Eickhoff, H., et al. (2003). A nonredundant human protein chip for antibody screening and serum profiling. Molecular & Cellular Proteomics, 2(12), 1342-1349. doi:10.1074/mcp.T300001-MCP200.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-8959-1 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-895A-0
Genre: Journal Article
Alternative Title : Mol. Cell. Proteomics

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 Creators:
Lueking, Angelika1, Author
Possling, Alexandra1, Author
Huber, Otmar, Author
Beveridge, Allan2, Author              
Horn, Martin, Author
Eickhoff, Holger, Author
Schuchardt, Johannes, Author
Lehrach, Hans2, Author              
Cahill, Dolores J.1, Author
Affiliations:
1Max Planck Society, ou_persistent13              
2Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              

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 Abstract: 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.

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Language(s): eng - English
 Dates: 2003-12
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: -
 Identifiers: eDoc: 174921
ISI: 000187250900009
DOI: 10.1074/mcp.T300001-MCP200
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Title: Molecular & Cellular Proteomics
  Alternative Title : Mol. Cell. Proteomics
Source Genre: Journal
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Pages: - Volume / Issue: 2 (12) Sequence Number: - Start / End Page: 1342 - 1349 Identifier: ISSN: 1535-9476