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.

中文翻译：

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农杆菌介导人源干扰素-α-2b和抗HER2抗体介导的免疫细胞因子瞬时表达的优化

摘要

优化农杆菌浸润溶液的组成是通过使用根癌农杆菌提高植物中靶蛋白瞬时表达效率的一种方法。我们使用实验设计来研究一些引入农杆菌浸润培养基中的化学添加剂对靶免疫细胞因子产量的影响。结果表明，乙酰丁香酮（600μM）和表面活性剂Pluronic-68（0.2％）显着提高了目标蛋白的产量；硫辛酸（0–5μM）和聚维酮（0–0.5 g / L）对该参数没有显着影响；浓度为25–50 mM的抗坏血酸会大大降低产量。使用本研究中开发的特定组成的农用浸润溶液，发现目标蛋白的产量超过500μg/ g，对于在植物中实际生产重组蛋白而言足够高。