Open AccessPlant cell packs: a scalable platform for recombinant protein production and metabolic engineering
Author(s) -
Rademacher Thomas,
Sack Markus,
Blessing Daniel,
Fischer Rainer,
Holland Tanja,
Buyel Johannes
Publication year - 2019
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/pbi.13081
Subject(s) - nicotiana benthamiana , biology , plant cell , agroinfiltration , nicotiana tabacum , metabolic engineering , computational biology , daucus carota , recombinant dna , microbiology and biotechnology , biochemistry , botany , enzyme , gene
Summary Industrial plant biotechnology applications include the production of sustainable fuels, complex metabolites and recombinant proteins, but process development can be impaired by a lack of reliable and scalable screening methods. Here, we describe a rapid and versatile expression system which involves the infusion of Agrobacterium tumefaciens into three‐dimensional, porous plant cell aggregates deprived of cultivation medium, which we have termed plant cell packs ( PCP s). This approach is compatible with different plant species such as Nicotiana tabacum BY 2, Nicotiana benthamiana or Daucus carota and 10‐times more effective than transient expression in liquid plant cell culture. We found that the expression of several proteins was similar in PCP s and intact plants, for example, 47 and 55 mg/kg for antibody 2G12 expressed in BY 2 PCP s and N. tabacum plants respectively. Additionally, the expression of specific enzymes can either increase the content of natural plant metabolites or be used to synthesize novel small molecules in the PCP s. The PCP method is currently scalable from a microtiter plate format suitable for high‐throughput screening to 150‐mL columns suitable for initial product preparation. It therefore combined the speed of transient expression in plants with the throughput of microbial screening systems. Plant cell packs therefore provide a convenient new platform for synthetic biology approaches, metabolic engineering and conventional recombinant protein expression techniques that require the multiplex analysis of several dozen up to hundreds of constructs for efficient product and process development.