title: Recombinant human albumin supports single cell cloning of CHO cells in chemically defined media.
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.
authors.affiliation: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology (AIBN), St. Lucia 4072, Australia.
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhu
authors.forename: Jong Wei
authors.initials: JW
authors.lastname: Wooh
authors.forename: Jeff Jia Cheng
authors.initials: JJ
authors.lastname: Hou
authors.forename: Benjamin S
authors.initials: BS
authors.lastname: Hughes
authors.forename: Peter P
authors.initials: PP
authors.lastname: Gray
authors.forename: Trent P
authors.initials: TP
authors.lastname: Munro
mesh_terms: D000418:Albumins / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D019976:Cloning, Organism; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D000418:Albumins; D011994:Recombinant Proteins