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

ID: 556523.0, MPI für molekulare Genetik / Department of Vertebrate Genomics
Semi-automated Magnetic Bead-Based Antibody Selection from Phage Display Libraries
Authors:Konthur, Zoltán; Wilde, Jeannine; Lim, Theam Soon
Place of Publication:Berlin et al
Date of Publication (YYYY-MM-DD):2010-05-07
Title of Series:Springer Protocols
Copyright:© 2010 Springer Science+Business Media
Audience:Experts Only
Abstract / Description:Phage display of combinatorial antibody libraries is a very efficient method for selecting recombinant antibodies against a wide range of molecules. It has been applied very successfully for the generation of therapeutic antibodies for more than a decade. To increase robustness and reproducibility of the selection procedure, we developed a semi-automated selection method for the generation of recombinant antibodies from phage display libraries. In this procedure, the selection targets are specifically immobilised to magnetic particles which can then by automatically handled by a magnetic particle processor. At present up to 96 samples can be handled simultaneously. Applying the processor allows standardisation of panning parameters such as washing conditions, incubation times, or to perform parallel selections on same targets under different buffer conditions. Additionally, the whole protocol has been streamlined to carry out bead loading, phage selection, phage amplification between selection rounds and magnetic particle ELISA for confirmation of binding activity in microtiter plate formats. Until now, this method has been successfully applied to select antibody fragments against different types of target, such as peptides, recombinant or homologous proteins, or chemical compounds.
External Publication Status:published
Document Type:InBook
Communicated by:Hans Lehrach
Affiliations:MPI für molekulare Genetik
External Affiliations:Faculty of Biology, Chemistry and Pharmacy, Free University Berlin, Takustrasse 3, Berlin, 14195, Germany
Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, 11800, Malaysia
Identifiers:ISBN:978-3-642-01143-6 [ID No:1]
DOI:10.1007/978-3-642-01144-3_18 [ID Nr:2]
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