Fraunhofer-Gesellschaft

Publica

Hier finden Sie wissenschaftliche Publikationen aus den Fraunhofer-Instituten.

Monoclonal tobacco cell lines with enhanced recombinant protein yields can be generated from heterogeneous cell suspension cultures by flow sorting

 
: Kirchhoff, Janina; Raven, Nicole; Boes, Alexander; Roberts, Jean L.; Russell, Sean; Treffenfeldt, Wiltrud; Fischer, Rainer; Schinkel, Helga; Schiermeyer, Andreas; Schillberg, Stefan

:

Plant biotechnology journal 10 (2012), No.8, pp.936-944
ISSN: 1467-7644 (print)
ISSN: 1467-7652 (online)
English
Journal Article
Fraunhofer IME ()
BY-2; cell line optimization; fluorescence-activated cell sorting; monoclonal human antibody; nicotiana tabacum; plant cell suspension culture; single-cell regeneration

Abstract
Plant cell suspension cultures can be used for the production of recombinant pharmaceutical proteins, but their potential is limited by modest production levels that may be unstable over long culture periods, reflecting initial culture heterogeneity and subsequent genetic and epigenetic changes. We used flow sorting to generate highly productive monoclonal cell lines from a heterogeneous population of tobacco BY-2 cells expressing the human antibody M12 by selecting the co-expressed ?uorescent marker protein DsRed located on the same T-DNA. Separation yielded ~35% wells containing single protoplasts and ~15% wells with monoclonal microcolonies that formed within 2 weeks. Thus, enriching the population of fluorescent cells from initially 24% to 90-96% in the six monoclonal lines resulted in an up to 13-fold increase in M12 production that remained stable for 10-12 months. This is the first straight-forward procedure allowing the generation of monoclonal plant cell suspension cultures by flow sorting, greatly increasing the potential of plant cells as an economical platform for the manufacture of recombinant pharmaceutical proteins.

: http://publica.fraunhofer.de/documents/N-213895.html