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Bachl, J. ; Caldwell, R.B. ; Buerstedde, J.M.

Biotechnology and the chicken B cell line DT40.

Cytogenet. Genome Res. 117, 189-194 (2007)
DOI PMC
Protein optimization is a major focus of the biotech and pharmaceutical industry. Various in vitro technologies have been developed to accelerate protein evolution and to achieve protein optimization of functional characteristics such as substrate specificity, enzymatic activity and thermostability. The chicken B cell line DT40 diversifies its immunoglobulin (Ig) gene by gene conversion and somatic hypermutation. This machinery can be directed to almost any gene inserted into the Ig locus. Enormously diverse protein libraries of any gene of interest can be quickly generated in DT40 by utilizing random shuffling of complex genetic domains (gene conversion) and by the introduction of novel non-templated genetic information (random mutagenesis). The unique characteristics of the chicken cell line DT40 make it a powerful in-cell diversification system to improve proteins of interest within living cells. One essential advantage of the DT40 protein optimization approach is the fact that variants are generated within an in-cell system thus allowing the direct screening for desired features in the context of intracellular networks. Utilizing specially designed selection strategies, such as the powerful fluorescent protein technology, enables the reliable identification of protein variants exhibiting the most desirable traits. Thus, DT40 is well positioned as a biotechnological tool to generate optimized proteins by applying a powerful combination of gene specific hypermutation, gene conversion and mutant selection.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Green fluorescent protein; Directed enzyme evolution; In-vitro recombination; Somatic hypermutation; Gene conversion; Phage display; Region hypermutation; Antibody diversity; Escherichia-coli; Lympocyte line
ISSN (print) / ISBN 0301-0171
e-ISSN 1421-9816
Journal Cytogenetic and Genome Research
Quellenangaben Volume: 117, Issue: 1-4, Pages: 189-194 Article Number: , Supplement: ,
Publisher Karger
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) Institute of Molecular Radiation Biology (IMS)