Development and Application of an Animalcomponent Free Single-cell Cloning Medium for Chinese Hamster Ovary Cell Lines View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2007

AUTHORS

N. Lin , J. L. Beckmann , J. Cresswell , J. S. Ross , B. Delong , Z. Deeds , M. V. Caple

ABSTRACT

Single-cell cloning is a critical process in the generation of recombinant protein-producing mammalian cell lines. This process traditionally requires 10 – 20% fetal bovine serum (FBS) or other sera. Due to the presence of sera, single-cell cloning is a potential source of contamination from animal viruses and other adventitious agents. In order to address the regulatory needs of the biopharmaceutical industry, we have developed an animal-component free (AF) medium designed for single-cell cloning of Chinese Hamster Ovary (CHO) cells. We utilized Design of Experiment (DOE) methodology to optimize the levels of six groups of nutrients (amino acids, trace metals, plantderived hydrolysates, lipids, vitamins and selenium). The optimized formulation was tested with three recombinant CHO cell lines and was shown to generate comparable results, in terms of clonal survival and growth (80% of positive control in average), to the 10% FBS control. The clones generated using the AF cloning medium from three recombinant CHO cell lines were successfully scaled up to spinner or shaker flask cultures in animal-component free culture media. The AF cloning process demonstrated improvement of growth and/or productivity. Transfection of a parental CHO K1 cell line and the subsequent selection and cloning processes were also evaluated in this AF cloning medium. More... »

PAGES

595-597

Book

TITLE

Cell Technology for Cell Products

ISBN

978-1-4020-5475-4

Author Affiliations

Patents

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-4020-5476-1_105

DOI

http://dx.doi.org/10.1007/978-1-4020-5476-1_105

DIMENSIONS

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