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Recombinant human albumin supports single cell cloning of CHO cells in chemically defined media
Author(s) -
Zhu Jiang,
Wooh Jong Wei,
Hou Jeff Jia Cheng,
Hughes Benjamin S.,
Gray Peter P.,
Munro Trent P.
Publication year - 2012
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.1549
Subject(s) - chinese hamster ovary cell , cloning (programming) , fetal bovine serum , cell culture , recombinant dna , chemically defined medium , bovine serum albumin , cell , monoclonal antibody , microbiology and biotechnology , biology , clone (java method) , chemistry , antibody , biochemistry , in vitro , immunology , genetics , gene , computer science , programming language
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