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Accelerated cell line development using two-color fluorescence activated cell sorting to select highly expressing antibody-producing clones.
Sleiman, R.J, Gray, Peter P., McCall, M.N., Giuseppe Codamo and Sunstrom, N-A, S. (2007-08-06) Accelerated cell line development using two-color fluorescence activated cell sorting to select highly expressing antibody-producing clones.. Biotechnology and Bioengineering, 99 3: 578-586.
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| Author(s) |
Sleiman, R.J
Gray, Peter P.
McCall, M.N.
Giuseppe Codamo
Sunstrom, N-A, S.
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| Title |
Accelerated cell line development using two-color fluorescence activated cell sorting to select highly expressing antibody-producing clones.
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| Journal name |
Biotechnology and Bioengineering
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| Publication date |
2007-08-06
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| Volume number |
99
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| Issue number |
3
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| ISSN |
0006-3592
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| Start page |
578
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| End page |
586
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| Place of publication |
United States of America
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| Publisher |
John Wiley & Sons, Inc.
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| Subject |
C1
100302 Bioprocessing, Bioproduction and Bioproducts
8608 Human Pharmaceutical Products
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| Abstract |
The success of engineered monoclonal antibodies as biopharmaceuticals has generated considerable interest in strategies designed to accelerate development of antibody expressing cell lines. Stable mammalian cell lines that express therapeutic antibodies at high levels typically take 6-12 months to develop. Here we describe a novel method to accelerate selection of cells expressing recombinant proteins (e.g., antibodies) using multiparameter fluorescence activated cell sorting (FACS) in association with dual intracellular autofluorescent reporter proteins. The method is co-factor-independent and does not require complex sample preparation. Chinese hamster ovary (CHO) clones expressing high levels of recombinant antibody were selected on the basis of a two-color FACS sorting strategy using heavy and light chain-specific fluorescent reporter proteins. We were able to establish within 12 weeks of transfection cell lines with greater than a 38-fold increase in antibody production when compared to the pool from which they were isolated, following a single round of FACS. The method provides a robust strategy to accelerate selection and characterization of clones and builds a foundation for a predictive model of specific productivity based upon on two-color fluorescence.
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| Additional Notes |
Published online 6 August 2007, then in hard copy Feb 15 2008. Should have been claimed in as 2007 publication for 08 HERDC
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