| Identification of Escherichia coli host cell for high plasmid stability and improved production of antihuman ovarian carcinoma x antihuman CD3 single-chain bispecific antibody. | |
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MedLine Citation:
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PMID: 17598107 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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To improve the plasmid stability during the production of antihuman ovarian carcinoma x antihuman CD3 single-chain bispecific antibody (AhOCxAhCD3), the Escherichia coli BL21(DE3) host cell was optimized serially. Firstly, an isogenic recombination-deficient (recA (-)) derivative of BL21(DE3), namely BLR(DE3), was used as host instead of BL21(DE3). Although the segregational plasmid stability was greatly improved, AhOCxAhCD3 yield was not improved due to the severe growth inhibition of plasmid-bearing BLR(DE3) cells and the competitive plasmid instability after induction. Secondly, a mutant BLR(DE3), namely BLRM(DE3), was screened by using LB agar plates plus ampicillin and isopropyl-beta-D: -thiogalactopyranoside. Using this new host, growth inhibition of recombinant cells after induction was eliminated, and plasmids could be stably maintained even after long-time induction in a nonselective medium. At last, about 1.2 g/l AhOCxAhCD3, which was about thrice as much as those of recombinant BL21(DE3) and BLR(DE3) strains, was yielded. |
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Authors:
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Jing-Bo Zhao; Dong-Zhi Wei; Wang-Yu Tong |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2007-06-28 |
Journal Detail:
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Title: Applied microbiology and biotechnology Volume: 76 ISSN: 0175-7598 ISO Abbreviation: Appl. Microbiol. Biotechnol. Publication Date: 2007 Sep |
Date Detail:
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Created Date: 2007-08-29 Completed Date: 2007-11-13 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 8406612 Medline TA: Appl Microbiol Biotechnol Country: Germany |
Other Details:
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Languages: eng Pagination: 795-800 Citation Subset: IM |
Affiliation:
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State Key Laboratory of Bioreactor Engineering, Newworld Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Antibodies, Bispecific
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biosynthesis*,
genetics,
immunology Antigens, CD3 / genetics, immunology* Biotechnology / methods* Escherichia coli / genetics Female Gene Deletion Humans Immunotherapy / methods Ovarian Neoplasms / immunology*, therapy Plasmids / genetics, immunology Rec A Recombinases / genetics Recombinant Proteins / biosynthesis, genetics, immunology |
| Chemical | |
Reg. No./Substance:
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0/Antibodies, Bispecific; 0/Antigens, CD3; 0/Recombinant Proteins; EC 2.7.7.-/Rec A Recombinases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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