Open AccessMonoclonal tobacco cell lines with enhanced recombinant protein yields can be generated from heterogeneous cell suspension cultures by flow sorting
Author(s) -
Kirchhoff Janina,
Raven Nicole,
Boes Alexander,
Roberts Jean L.,
Russell Sean,
Treffenfeldt Wiltrud,
Fischer Rainer,
Schinkel Helga,
Schiermeyer Andreas,
Schillberg Stefan
Publication year - 2012
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/j.1467-7652.2012.00722.x
Subject(s) - monoclonal antibody , biology , cell sorting , flow cytometry , recombinant dna , cell culture , population , green fluorescent protein , monoclonal , suspension culture , sorting , protoplast , microbiology and biotechnology , biochemistry , antibody , genetics , gene , demography , sociology , computer science , programming language
Summary 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 fluorescent 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 straightforward 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.