Recombinant human albumin supports single cell cloning of CHO cells in chemically defined media

Authors

  • Jiang Zhu,

    1. The University of Queensland, Australian Institute for Bioengineering and Nanotechnology (AIBN), St. Lucia 4072, Australia
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  • Jong Wei Wooh,

    1. The University of Queensland, Australian Institute for Bioengineering and Nanotechnology (AIBN), St. Lucia 4072, Australia
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  • Jeff Jia Cheng Hou,

    1. The University of Queensland, Australian Institute for Bioengineering and Nanotechnology (AIBN), St. Lucia 4072, Australia
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  • Benjamin S. Hughes,

    1. The University of Queensland, Australian Institute for Bioengineering and Nanotechnology (AIBN), St. Lucia 4072, Australia
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  • Peter P. Gray,

    1. The University of Queensland, Australian Institute for Bioengineering and Nanotechnology (AIBN), St. Lucia 4072, Australia
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  • Trent P. Munro

    Corresponding author
    1. The University of Queensland, Australian Institute for Bioengineering and Nanotechnology (AIBN), St. Lucia 4072, Australia
    • Australian Institute for Bioengineering and Nanotechnology (AIBN), The University of Queensland, St. Lucia 4072, Australia
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Abstract

Biologic drugs, such as monoclonal antibodies, are commonly made using mammalian cells in culture. The cell lines used for manufacturing should ideally be clonal, meaning derived from a single cell, which represents a technically challenging process. Fetal bovine serum is often used to support low cell density cultures, however, from a regulatory perspective, it is preferable to avoid animal-derived components to increase process consistency and reduce the risk of contamination from adventitious agents. Chinese hamster ovary (CHO) cells are the most widely used cell line in industry and a large number of serum-free, protein-free, and fully chemically defined growth media are commercially available, although these media alone do not readily support efficient single cell cloning. In this work, we have developed a simple, fully defined, single-cell cloning media, specifically for CHO cells, using commercially available reagents. Our results show that a 1:1 mixture of CD-CHO and DMEM/F12 supplemented with 1.5 g/L of recombinant albumin (Albucult®) supports single cell cloning. This formulation can support recovery of single cells in 43% of cultures compared to 62% in the presence of serum. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012

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