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          Institute: MPI für molekulare Genetik     Collection: Department of Vertebrate Genomics     Display Documents

ID: 229481.0, MPI für molekulare Genetik / Department of Vertebrate Genomics
Seeing better through a MIST : evaluation of monoclonal recombinant antibody fragments on microarrays
Authors:Angenendt, Philipp; Wilde, Jeannine; Kijanka, Gregor; Baars, Sabine; Cahill, Dolores J.; Kreutzberger, Jürgen; Lehrach, Hans; Konthur, Zoltán; Glökler, Jörn
Date of Publication (YYYY-MM-DD):2004-10
Title of Journal:Analytical Chemistry
Journal Abbrev.:Anal Chem
Issue / Number:10
Start Page:2916
End Page:2921
Copyright:© 2004 American Chemical Society
Review Status:not specified
Audience:Experts Only
Abstract / Description:Automation is the key approach for genomewide and proteomewide screening of function and interaction. Especially for proteomics, antibody microarrays are a useful tool for massive parallel profiling of complex samples. To meet the requirements of antibody microarrays and to obtain a great variety of antibodies, new technologies such as phage display have partly replaced the classical hybridoma method. While the selection process for phage-displayed antibody fragments itself has been automated, the bottleneck was shifted further downstream to the identification of monoclonal binders obtained from the selections. Here, we present a new approach to reduce time, material, and waste to extend automation beyond the selection process by application of conventional microarray machinery. We were able to express recombinant antibody fragments in a single inoculation and expression step and subjected them without purification directly to an automated high-throughput screening procedure based on the multiple spotting technique (MIST). While obtaining comparable sensitivities to enzyme-linked immunosorbent assays, we minimized manual interaction steps and streamlined the technique to be accessible within the automated selection procedure.
Comment of the Author/Creator:Published online: 2004-04-13
External Publication Status:published
Document Type:Article
Communicated by:Hans Lehrach
Affiliations:MPI für molekulare Genetik
External Affiliations:Medical Proteome Center, Ruhr-University Bochum, Universitätsstrasse 150, 44780 Bochum, Germany;
PROTAGEN AG, Emil-Figge-Strasse 76A, 44227 Dortmund, Germany; The Centre for Human Proteomics, Royal College of Surgeons in Ireland, Dublin 2, Ireland
DOI:10.1021/ac035357a S0003-2700(03)05357-5
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