A nonredundant human protein chip for antibody screening and serum profiling
| Autor: | Angelika Lueking, Martin Horn, Alexandra Possling, Allan Beveridge, Hans Lehrach, Holger Eickhoff, Dolores J. Cahill, Johannes Schuchardt, Otmar Huber |
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| Předmět: |
Proteome
Microscope slide Protein Array Analysis Plasma protein binding Biology Cross Reactions Biochemistry Antibodies Analytical Chemistry Protein–protein interaction Arthritis Rheumatoid Mice Antibody Repertoire Antibody Specificity Animals Humans Molecular Biology Gene Library Immune Sera Alopecia Molecular biology Fusion protein Protein microarray Protein Binding |
| Zdroj: | Scopus-Elsevier |
| Popis: | 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/cm(2) 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-microl 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-RGSHis(6), anti-GAPDH, and anti-HSP90beta 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. |
| Databáze: | OpenAIRE |
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