Translation initiation is on the critical pathway for the production of monoclonal antibodies (mAbs) by mammalian cells. Formation of a closed loop structure comprised of mRNA, a number of eukaryotic initiation factors (eIFs) and ribosomal proteins has been proposed to aid re-initiation of translation and therefore increase global translational efficiency. We have determined mRNA and protein levels of the key components of the closed loop, eIFs (eIF3a, eIF3b, eIF3c, eIF3h, eIF3i and eIF4G1), poly(A)-binding protein (PABP) 1 and PABP-interacting protein 1 (PAIP1), across a panel of 30 recombinant mAb-producing GS-CHOK1SV cell lines with a broad range of growth characteristics and production levels of a model recombinant mAb. We have used a multi-level statistical approach to investigate the relationship between key performance indicators (cell growth and recombinant antibody productivity) and the intracellular amounts of target translation initiation factor proteins and the mRNAs encoding them. We show that high-producing cell lines maintain amounts of the translation initiation factors involved in the formation of the closed loop mRNA, maintaining these proteins at appropriate levels to deliver enhanced recombinant protein production. We then utilize knowledge of the amounts of these factors to build predictive models for and use cluster analysis to identify, high-producing cell lines. The present study therefore defines the translation initiation factor amounts that are associated with highly productive recombinant GS-CHOK1SV cell lines that may be targets for screening highly productive cell lines or to engineer new host cell lines with the potential for enhanced recombinant antibody productivity.
Skip Nav Destination
Article navigation
December 2015
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
November 27 2015
Biological insights into the expression of translation initiation factors from recombinant CHOK1SV cell lines and their relationship to enhanced productivity
Emma J. Mead;
Emma J. Mead
1
*Centre for Molecular Processing and School of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, U.K.
1Correspondence may be addressed to either of these authors (email e.j.hargreaves@kent.ac.uk or c.m.smales@kent.ac.uk).
Search for other works by this author on:
Rosalyn J. Masterton;
Rosalyn J. Masterton
*Centre for Molecular Processing and School of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, U.K.
Search for other works by this author on:
Marc Feary;
Marc Feary
†Lonza Biologics plc, Portway Building, Granta Park, Great Abington, Cambridge CB21 6GS, U.K.
Search for other works by this author on:
Olga Obrezanova;
Olga Obrezanova
†Lonza Biologics plc, Portway Building, Granta Park, Great Abington, Cambridge CB21 6GS, U.K.
Search for other works by this author on:
Lin Zhang;
Lin Zhang
‡Pfizer Inc, 1 Burtt Road, Andover, MA 01810, U.S.A.
Search for other works by this author on:
Robert Young;
Robert Young
†Lonza Biologics plc, Portway Building, Granta Park, Great Abington, Cambridge CB21 6GS, U.K.
Search for other works by this author on:
C. Mark Smales
C. Mark Smales
1
*Centre for Molecular Processing and School of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, U.K.
1Correspondence may be addressed to either of these authors (email e.j.hargreaves@kent.ac.uk or c.m.smales@kent.ac.uk).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
August 21 2015
Revision Received:
September 28 2015
Accepted:
September 29 2015
Accepted Manuscript online:
September 29 2015
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© 2015 Authors; published by Portland Press Limited
2015
Biochem J (2015) 472 (3): 261–273.
Article history
Received:
August 21 2015
Revision Received:
September 28 2015
Accepted:
September 29 2015
Accepted Manuscript online:
September 29 2015
Connected Content
Citation
Emma J. Mead, Rosalyn J. Masterton, Marc Feary, Olga Obrezanova, Lin Zhang, Robert Young, C. Mark Smales; Biological insights into the expression of translation initiation factors from recombinant CHOK1SV cell lines and their relationship to enhanced productivity. Biochem J 15 December 2015; 472 (3): 261–273. doi: https://doi.org/10.1042/BJ20150928
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.