alexa Deep sequencing reveals different compositions of mRNA transcribed from the F8 gene in a panel of FVIII-producing CHO cell lines.
Chemical Engineering

Chemical Engineering

Journal of Bioprocessing & Biotechniques

Author(s): Kaas CS, , Bolt G, Hansen JJ, Andersen MR,

Abstract Share this page

Abstract Coagulation factor VIII (FVIII) is one of the most complex biopharmaceuticals due to the large size, poor protein stability and extensive post-translational modifications. As a consequence, efficient production of FVIII in mammalian cells poses a major challenge, with typical yields two to three orders of magnitude lower than for antibodies. In the present study we investigated CHO DXB11 cells transfected with a plasmid encoding human coagulation factor VIII. Single cell clones were isolated from the pool of transfectants and a panel of 14 clones representing a dynamic range of FVIII productivities was selected for RNA sequencing analysis. The analysis showed distinct differences in F8 RNA composition between the clones. The exogenous F8-dhfr transcript was found to make up the most abundant transcript in the present clones. No correlation was seen between F8 mRNA levels and the measured FVIII productivity. It was found that three MTX resistant, nonproducing clones had different truncations of the F8 transcripts. We find that by using deep sequencing, in contrast to microarray technology, for determining the transcriptome from CHO transfectants, we are able to accurately deduce the mature mRNA composition of the transgene and identify significant truncations that would probably otherwise have remained undetected. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. This article was published in Biotechnol J and referenced in Journal of Bioprocessing & Biotechniques

Relevant Expert PPTs

Relevant Speaker PPTs

Recommended Conferences

  • 17th Euro Biotechnology Congress
    September 25-27, 2017 Berlin, Germany
  • 2nd World Biotechnology Congress
    December 04-06, 2017 Sao Paulo, Brazil
Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2017-18
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

 
© 2008-2017 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version
adwords