Skip to main content
Cytotechnology logoLink to Cytotechnology
. 2002 Jan;38(1-3):43–46. doi: 10.1023/A:1021141712344

The use of UCOE vectors in combination with a preadapted serum free, suspension cell line allows for rapid production of large quantities of protein

Trish Benton 1, Tim Chen 2, Michele McEntee 2, Brian Fox 2, David King 2, Robert Crombie 3, Thomas C Thomas 2, Christopher Bebbington 2
PMCID: PMC3449923  PMID: 19003085

Abstract

UCOE vectors contain non-tissue specific chromatin-opening-elements that permit rapid expression of a protein in anintegration independent manner. Efficient expression can bederived from a single copy of an integrated gene site resulting ina higher percentage of cells expressing the marker gene in theselected pool in comparison to standard non-UCOE containingvectors. This, in combination with the utilization of a serum-free, suspension adapted parent cell line allows for rapidproduction of large quantities of protein in a short period oftime. Utilizing this system more than 300 mg of a recombinantantibody has been produced in less than 1 month from transfectionpools in shake flask. Selected subclones have been scaled intosmall bioreactors in less than 2 months, producing significantquantities of monoclonal antibody using a protocol generic for theparent cell line. The increased efficiency obtained with the UCOEvector reduces the number of transfectants which need to bescreened in order to obtain high productivity subclones.Transfection of a standard host cell line, preadapted to grow in alarge-scale setting, allows for rapid cell line developmentdecreasing the transition time from research into development andmanufacturing. Alternatively, the traditional approach of using aparent cell line which requires serum-free and suspensionadaptation after transfection further increases the need forscreening a large number of subclones, because many of thesubclones will not be able to grow under conditions that allowlarge-scale protein production. The use of a preadapted cell linecan reduce the time required to develop a cell line from months toweeks.

Keywords: preadapted serum-free cell culture, rapid proteinproduction, UCOE

Full Text

The Full Text of this article is available as a PDF (80.5 KB).

References

  1. Borrebeack CM, Ohlin M. Does endogenous glycosylation prevent the use of mouse monoclonal antibodies as cancer therapeutics? Immunol Today. 1993;14:477–479. doi: 10.1016/0167-5699(93)90259-n. [DOI] [PubMed] [Google Scholar]
  2. Cho SK, Cummings RD. A soluble form of α1,3-galactosyltransferase functions within cells to glycosylate proteins. J Biol Chem. 1997;272:13622–13628. doi: 10.1074/jbc.272.21.13622. [DOI] [PubMed] [Google Scholar]
  3. Crombie R & Antoniou M (2000) A Polynucleotide Comprising a Ubiquitous Chromatin Opening Element (UCOE). Patent Number WO0005393, 2000-02-03.
  4. D'Anna JA, Valdez JG, Habbersett RC, Crissman HA. Association of G1/S-Phase and Late S-Phase Checkpoints with Regulation of Cyclin-Dependent Kinases in Chinese Hamster Ovary Cells. Rad Res. 1997;148:260–271. [PubMed] [Google Scholar]
  5. D'Anna JA, Crissman HA, Valdez JG, Gurley LR, Tobey RA. Syncronization of mammalian cells in S-phase by sequential use of isoleucine-deprivation G1-or serum-withdrawl G0-arrest and aphidicolin block. Meth Cell Sci. 1996;18:115–125. [Google Scholar]
  6. Deaven LL, Petersen DF. The Chromosomes of CHO, an Aneuploid Chinese Hamster Cell Line: G-Band, C-Band, and Autoradiographic Analysis. Chromosoma. 1973;41:129–144. doi: 10.1007/BF00319690. [DOI] [PubMed] [Google Scholar]
  7. Gorelik EL, Anaraki DF, Galili U. Alterations of cell surface carbohydrate and inhibition of metastatic property of murine melanomas by α1,3 galactosyltransferase gene transfection. Cancer Res. 1995;55:4185–4173. [PubMed] [Google Scholar]
  8. Gorfien SF, Dzimian JL, Tilkins ML, Godwin GP, Fike R. Recombinant Protein Production by CHO Cells Cultures in a Chemically Defined Medium. In: Kitagawa Y, Matsuda T, Iijima S, editors. Animal Cell Technology: Basic & Applied Aspects. (Netherlands): Kluwer Academic Publishers; 1998. pp. 247–252. [Google Scholar]
  9. Kagawa YT, Utsumi S, Hosoi K, Shimizu H, Kochibe N, Kobata A. Comparative study of the asparagines-linked sugar chains of natural human interferon-1 and recombinant human interferon produced by three different mammalian cell lines. J Biol Chem. 1988;263:7508–17515. [PubMed] [Google Scholar]
  10. Puck TT, Cieciura SJ, Robinson A. Genetics of somatic mammalian cells: Long term cultivation of euploid cells from human and animal subjects. J Exp Med. 1958;108:945–955. doi: 10.1084/jem.108.6.945. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Cytotechnology are provided here courtesy of Springer Science+Business Media B.V.

RESOURCES