Macharoen Kantharakorn, McDonald Karen A, Nandi Somen
Department of Chemical Engineering, University of California, Davis, CA 95616, USA.
Global HealthShare® Initiative, University of California, Davis, CA 95616, USA.
MethodsX. 2020 Nov 12;7:101139. doi: 10.1016/j.mex.2020.101139. eCollection 2020.
Transgenic plant cell suspension culture is a promising platform for recombinant protein production. Rice cell suspension culture is one of the systems that has been developed due to its unique metabolically-regulated promoter, rice alpha-amylase 3D (), that is up-regulated in sugar-deprived medium. Using the promoter system in transgenic rice cell suspensions results in two phases of the culture, the growth phase and the induction phase. Conventionally, medium exchange is performed to remove residual sugar and induce recombinant protein. In this work, a simplified production process is demonstrated in a 5-L bioreactor, including reduction of sugar concentration in the initial culture medium, elimination of the media exchange operation, and uncontrolled dissolved oxygen (DO) with constant aeration. The simplified method significantly improves the accumulation level of a recombinant protein, protein purity, and productivity compared to the conventional method. This method also reduces costs associated with material and labor. • The method of simplified bioreactor processing includes single-stage culture, uncontrolled dissolved oxygen (DO) but controlled inlet air flowrate, and lower (50% reduction) initial sucrose concentration in the culture medium. • This method improves recombinant protein production level and productivity compared to the conventional method. • This method reduces material and labor costs.
转基因植物细胞悬浮培养是一种很有前景的重组蛋白生产平台。水稻细胞悬浮培养是已开发的系统之一，因其具有独特的代谢调控启动子——水稻α淀粉酶3D（），该启动子在无糖培养基中会上调。在转基因水稻细胞悬浮液中使用该启动子系统会导致培养分为两个阶段，即生长阶段和诱导阶段。传统上，通过更换培养基来去除残留糖分并诱导重组蛋白表达。在这项研究中，在5升生物反应器中展示了一种简化的生产工艺，包括降低初始培养基中的糖浓度、取消培养基更换操作以及在恒定通气条件下不控制溶解氧（DO）。与传统方法相比，该简化方法显著提高了重组蛋白的积累水平、蛋白纯度和生产率。该方法还降低了材料和劳动力成本。• 简化生物反应器处理方法包括单阶段培养、不控制溶解氧（DO）但控制进气流量，以及降低（降低50%）培养基中的初始蔗糖浓度。• 与传统方法相比，该方法提高了重组蛋白的生产水平和生产率。• 该方法降低了材料和劳动力成本。
