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Detection of cytosine deaminase in genetically modified tumor cells by specific antibodies

Bacterial cytosine deaminase (CD) converts the non-toxic prodrug 5-fluorocytosine (5-FC) into 5-fluorouracil (5-FU), which is toxic for mammalian cells. Therefore, the CD gene is used in cancer gene therapy to achieve high local concentration of a toxic metabolite without significant systemic toxici...

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Bibliographic Details
Main Authors: Haack, Karin (Author) , Moebius, Ulrich (Author) , Knebel Doeberitz, Magnus von (Author) , Herfarth, Christian (Author) , Schackert, Hans K. (Author) , Gebert, Johannes (Author)
Format: Article (Journal)
Language:English
Published: 1997
In: Human gene therapy
Year: 1997, Volume: 8, Issue: 11, Pages: 1395-1401
ISSN:1557-7422
DOI:10.1089/hum.1997.8.11-1395
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1089/hum.1997.8.11-1395
Verlag, lizenzpflichtig, Volltext: https://www.liebertpub.com/doi/10.1089/hum.1997.8.11-1395
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Author Notes:K. Haack, U. Moebius, M.V. Knebel Doeberitz, Ch. Herfarth, H.-K. Schackert, J.F. Gebert
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Summary:Bacterial cytosine deaminase (CD) converts the non-toxic prodrug 5-fluorocytosine (5-FC) into 5-fluorouracil (5-FU), which is toxic for mammalian cells. Therefore, the CD gene is used in cancer gene therapy to achieve high local concentration of a toxic metabolite without significant systemic toxicity. To allow the detection of CD expression at the protein level, we raised both polyclonal rabbit antisera and a monoclonal antibody (mAb) against a histidine-tagged CD fusion protein. The specificity of the polyclonal antisera and the mAb was confirmed by immunohistochemistry, immunoblot analysis, and immunoprecipitation using CD-expressing tumor cell lines. Furthermore, the antibodies can be used for ELISA assays and flow cytometry. Finally, the CD protein could be demonstrated in frozen tissue sections of CD-modified tumors in a rat tumor model using the anti-CD serum. With these antibodies, CD expression can now be monitored throughout in vitro and in vivo gene transfer studies, including clinical protocols relying on the CD suicide gene strategy.
Item Description:Elektronische Reproduktion der Druck-Ausgabe 20. März 2008
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Physical Description:Online Resource
ISSN:1557-7422
DOI:10.1089/hum.1997.8.11-1395

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