Process development of cytokines production for regenerative medicines

by transient expression in Nicotiana benthamiana

Yasumasa Dekishima

Medical Core Project Division, Innovation Strategy Department,

Mitsubishi Chemical Corporation

Cytokines are a group of proteins playing very important roles in the manufacture of regenerative medicine and relating products, using iPSC (pluripotent stem cells)/ESC (embryonic stem cells) or MSC (mesenchymal stem cells), to promote their proliferation or differentiation in cell culture media. Although recombinant protein production system such as mammalian cells, bacteria, yeast, and insect cells have been developed, each system has its own merits and demerits in terms of production method, activity, and raw material/impurity control.

Under these circumstances, we have focused on the transient expression system using plants (Nicotiana benthamiana) and are working to apply this recombinant protein expression technology as a cytokines production. One of the advantages of using plant is that it does not use animal-derived raw materials and thus avoids contamination with viruses that may pose a risk of infection to humans. It is also known that proteins that aggregate in bacteria such as Escherichia coli could be expressed as soluble manner in plants by controlling the location of expression.

On the other hand, host development (protease-deficient strains, molecular chaperones, etc.) suitable for heterologous protein expression, as has been done in E. coli, has not been studied well in plants, and degradation of recombinant proteins because of endogenous proteases becomes problem.

In the presentation, we will describe our recent work for producing cytokines by transient expression in N. benthamiana using Activin A, a member of the transforming growth factor β superfamily, as an example.

Reference:

WO 2020100933, JP 2020-156412