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Fluorescent labeling in semi-solid medium for selection of mammalian cells secreting high-levels of recombinant proteins |
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| Authors: | Antoine W Caron Claire Nicolas Bruno Gaillet Ismaïla Ba Maxime Pinard Alain Garnier Bernard Massie Rénald Gilbert |
| Institution: | 1. Institut de Recherche en Biotechnologie, Conseil National de Recherches du Canada, 6100 Royalmount Avenue, Montréal, QC H4P 2R2, Canada 2. Chemical Engineering Department, Université Laval, Québec, QC, G1K 7P4, Canada 3. Département de Microbiologie et Immunologie, Faculté de Médecine, Université de Montréal, Montreal, QC, H3C 3J7, Canada 4. INRS-IAF, Université du Québec, Laval, QC, H7N 4Z3, Canada 5. Neuromuscular research group, Montreal Neurological Institute, Montreal, QC, H3A 2B4, Canada |
| Abstract: | BackgroundDespite the powerful impact in recent years of gene expression markers like the green fluorescent protein (GFP) to link the expression of recombinant protein for selection of high producers, there is a strong incentive to develop rapid and efficient methods for isolating mammalian cell clones secreting high levels of marker-free recombinant proteins. Recently, a method combining cell colony growth in methylcellulose-based medium with detection by a fluorescently labeled secondary antibody or antigen has shown promise for the selection of Chinese Hamster Ovary (CHO) cell lines secreting recombinant antibodies. Here we report an extension of this method referred to as fluorescent labeling in semi-solid medium (FLSSM) to detect recombinant proteins significantly smaller than antibodies, such as IGF-E5, a 25 kDa insulin-like growth factor derivative. |
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