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K. M. Konoplina, E. N. Kosobokova, E. V. Sheshukova, M. V. Pinyugina, A. A. Malchenkova, V. S. Kosorukov
Optimization
of Agrobacterium-mediated transient expression of immunocytokine based
on human interferon-α-2b and anti-HER2 antibody in Nicotiana bentham
Abstract
Optimization
of the agroinfiltration media composition is one of the options to increase the
efficiency of the Agrobacterium tumefaciens-mediated transient
expression of the target protein in plants. In the study, we have demonstrated
effects of some chemical additives in the agroinfiltration media on the target
immunocytokine yield by statistical design of the experiment. It was shown that
acetosyringone (600 μM) and pluronic F-68 (0.2%) increased the target protein yield
significantly, while lipoic acid (0–5 μM) and povidone (0–0.5 g/L) did not have a significant
impact on the protein yield and ascorbic acid (25–50 mM) decreased the product
yield significantly. As a result of the study, the created composition of the
agroinfiltration media made it possible to produce 500 μg/g of the target protein yield, which
is a sufficient technological level for large-scale production of recombinant
proteins in plants.
Key words:
agroinfiltration, protein expression optimization, statistical design of the
experiment, transient expression, Nicotiana
benthamiana.
Copyright (C) Chemistry Dept., Moscow State University, 2002
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