Optimization of Agrobacterium-Mediated Transient Expression of Immunocytokine Based on Human Interferon-α-2b and Anti-HER2 Antibody in Nicotiana Benthamiana

Abstract

Optimizing the composition of an agroinfiltration solution is one way to boost the efficiency of the transient expression of the target protein in plants by using Agrobacterium tumefaciens. We use the design of the experiment to investigate the effects that some chemical additives introduced into the agroinfiltration medium have on the yield of the target immunocytokine. It is shown that acetosyringone (at 600 μM) and the surfactant Pluronic-68 (at 0.2%) markedly increase the yield of the target protein; lipoic acid (0–5 μM) and povidone (0–0.5 g/L) do not have a significant effect on this parameter; and ascorbic acid at concentrations of 25–50 mM significantly lowers the yield. Using an agroinfiltration solution with the specific composition developed in this study, the yield of the target protein is found to be more than 500 μg/g, which is sufficiently high for the practical production of recombinant proteins in plants.