Comprehensive characterization of glutamine synthetase-mediated selection for the establishment of recombinant CHO cells producing monoclonal antibodies

Cited 3 time in webofscience Cited 0 time in scopus
  • Hit : 126
  • Download : 41
To characterize a glutamine synthetase (GS)-based selection system, monoclonal antibody (mAb) producing recombinant CHO cell clones were generated by a single round of selection at various methionine sulfoximine (MSX) concentrations (0, 25, and 50 mu M) using two different host cell lines (CHO-K1 and GS-knockout CHO). Regardless of the host cell lines used, the clones selected at 50 mu M MSX had the lowest average specific growth rate and the highest average specific production rates of toxic metabolic wastes, lactate and ammonia. Unlike CHO-K1, high producing clones could be generated in the absence of MSX using GS-knockout CHO with an improved selection stringency. Regardless of the host cell lines used, the clones selected at various MSX concentrations showed no significant difference in the GS, heavy chain, and light chain gene copies (P > 0.05). Furthermore, there was no correlation between the specific mAb productivity and these three gene copies (R-2 <= 0.012). Taken together, GS-mediated gene amplification does not occur in a single round of selection at a MSX concentration up to 50 mu M. The use of the GS-knockout CHO host cell line facilitates the rapid generation of high producing clones with reduced production of lactate and ammonia in the absence of MSX.
Publisher
NATURE PUBLISHING GROUP
Issue Date
2018-03
Language
English
Article Type
Article
Keywords

HAMSTER OVARY CELLS; HIGH-LEVEL EXPRESSION; MESSENGER-RNA LEVEL; GENE COPY NUMBER; LINE GENERATION; MAMMALIAN-CELLS; MYELOMA CELLS; AMPLIFICATION; STABILITY; AMMONIA

Citation

SCIENTIFIC REPORTS, v.8, no.5361, pp.1 - 11

ISSN
2045-2322
DOI
10.1038/s41598-018-23720-9
URI
http://hdl.handle.net/10203/241443
Appears in Collection
BS-Journal Papers(저널논문)
Files in This Item
s41598-018-23720-9.pdf(2.11 MB)Download
This item is cited by other documents in WoS
⊙ Detail Information in WoSⓡ Click to see webofscience_button
⊙ Cited 3 items in WoS Click to see citing articles in records_button

qr_code

  • mendeley

    citeulike


rss_1.0 rss_2.0 atom_1.0