Accelerated cell line development using two-color fluorescence activated cell sorting to select highly expressing antibody-producing clones

Sleiman, Robert J., Gray, Peter P., McCall, Martin N., Codamo, Joe and Sunstrom, Noelle-Ann S. (2008) 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.


Author Sleiman, Robert J.
Gray, Peter P.
McCall, Martin N.
Codamo, Joe
Sunstrom, Noelle-Ann S.
Title Accelerated cell line development using two-color fluorescence activated cell sorting to select highly expressing antibody-producing clones
Journal name Biotechnology and Bioengineering  (ERA 2012 Listed)    (ERA 2010 Rank A)   Check publisher's open access policy
Publication date 2008-02-15
Sub-type Article
DOI 10.1002/bit.21612
Volume number 99
Issue number 3
ISSN 0006-3592
Start page 578
End page 586
Total pages 9
Place of publication Hoboken, N. J., U.S.
Publisher John Wiley & Sons, Inc.
Language eng
Subject C1
100302 Bioprocessing, Bioproduction and Bioproducts
8608 Human Pharmaceutical Products
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.
Keyword CHO
fluorescence activated cell sorting
GFP
selection
recombinant antibody
Q-Index Code C1
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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