Please use this identifier to cite or link to this item:
Title: Vector fragmentation: Characterizing vector integrity in transfected clones by Southern blotting
Authors: Ng, S.K.
Lin, W.
Sachdeva, R.
Wang, D.I.C.
Yap, M.G.S. 
Keywords: CHO
Dihydrofolate reductase
Integration site
Mammalian cell culture
Southern blot
Vector fragmentation
Vector integrity
Issue Date: Jan-2010
Citation: Ng, S.K., Lin, W., Sachdeva, R., Wang, D.I.C., Yap, M.G.S. (2010-01). Vector fragmentation: Characterizing vector integrity in transfected clones by Southern blotting. Biotechnology Progress 26 (1) : 11-20. ScholarBank@NUS Repository.
Abstract: The Chinese Hamster Ovary production cell line development process using methotrexate (MTX) amplification is well studied and commonly used for biopharmaceutical processes. However, successful MTX amplification varies from clone to clone and suggested reasons include vector fragmentation during the transfection process and genomic rearrangement of the Chinese Hamster Ovary chromosomes. Here, we elucidated the vector integration patterns of 40 transfected single-cell clones by Southern blotting and showed that vector fragmentation occurs at a significant level in our experiment. This concurs with MTX amplification studies implying that single-cell cloning is necessary to ensure a successful amplification process. Truncations at the ends of the integrated vectors were also observed, whereas gross DNA insertions were not detected in our data. This suggests that end deletions are common, whereas insertion events are rare in animal cells. © 2009 American Institute of Chemical Engineers.
Source Title: Biotechnology Progress
ISSN: 87567938
DOI: 10.1002/btpr.281
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.


checked on Mar 20, 2019


checked on Mar 11, 2019

Page view(s)

checked on Mar 16, 2019

Google ScholarTM



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.