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

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 c...

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Bibliographic Details
Published in:Biotechnology progress 2012-05, Vol.28 (3), p.887-891
Main Authors: Zhu, Jiang, Wooh, Jong Wei, Hou, Jeff Jia Cheng, Hughes, Benjamin S., Gray, Peter P., Munro, Trent P.
Format: Article
Language:English
Subjects:
Albumins - metabolism
Animals
Biological and medical sciences
Biotechnology
CHO Cells
Cloning, Organism
Cricetinae
Cricetulus
Fundamental and applied biological sciences. Psychology
Humans
recombinant albumin
Recombinant Proteins - metabolism
single-cell cloning
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Summary: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
ISSN:8756-7938
1520-6033
DOI:10.1002/btpr.1549