The Powerful Combination of Phage Surface Display of cDNA Libraries and High Throughput Screening

Authors: Crameri R.; Kodzius R.

Source: Combinatorial Chemistry & High Throughput Screening, Volume 4, Number 2, April 2001 , pp. 145-155(11)

Publisher: Bentham Science Publishers

Abstract:

Phage surface display of cDNA libraries facilitates cloning, expression and rapid selection of functional gene products physically linked to their genetic information through gene product-ligand interactions. Efficient screening technologies based on selective enrichment of clones expressing desired gene products allows, within a short time, the isolation of all ligand-specific clones that are present in a library. Manual identification of clones by restriction analysis and random sequencing is unlike to be successful for the isolation of gene products derived from rare mRNA species resulting from selection of the libraries using polyvalent ligands like serum from patients. Here we describe rapid handling of large numbers of individual clones selected from molecular libraries displayed on phage surface using the power of robotics-based high throughput screening. The potential of the combination of cDNA-phage surface display, with selection for specific interactions by functional screening and robotic technology is illustrated by the isolation of more sequences potentially encoding IgE-binding proteins than postulated from Western blot analyses using extracts derived from raw material of complex allergenic sources. The subsequent application of functional enrichment and robotics-based screening will facilitate the rapid generation of information about the repertoire of protein structures involved in allergic diseases.

Keywords: Phage Surface Display of cDNA Libraries; High Throughput Screening; Alternaria alternata; Aspergillus fumigatus; Candida albicans; Cladosporium herbarum; Coprinus comatus; Malassezia furfur; Malassezia furfur; Peanut; Storage mites

Language: English

Document Type: Review article

DOI: http://dx.doi.org/10.2174/1386207013331237

Publication date: 2001-04-01

• Combinatorial Chemistry & High Throughput Screening publishes full length original research articles and reviews describing various topics in combinatorial chemistry (e.g. small molecules, peptide, nucleic acid or phage display libraries) and/or high throughput screening (e.g. developmental, practical or theoretical). Ancillary subjects of key importance, such as robotics and informatics, will also be covered by the journal. In these respective subject areas, Combinatorial Chemistry & High Throughput Screening is intended to function as the most comprehensive and up-to-date medium available. The journal should be of value to individuals engaged in the process of drug discoveryand development, in the settings of industry, academia or government.

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• By this author: Crameri R. ;  Kodzius R.

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