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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| Main Authors: | , , , , , |
|---|---|
| Format: | Article (Journal) |
| Language: | English |
| Published: |
1997
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| 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 |
| Author Notes: | K. Haack, U. Moebius, M.V. Knebel Doeberitz, Ch. Herfarth, H.-K. Schackert, J.F. Gebert |
MARC
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| 520 | |a 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. | ||
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