Department of Chemical Engineering, University of California, Davis, CA 95616, USA.
Department of Chemistry, University of California, Davis, CA 95616, USA.
Int J Mol Sci. 2018 Apr 16;19(4):1205. doi: 10.3390/ijms19041205.
Transient recombinant protein production is a promising alternative to stable transgenic systems, particularly for emergency situations in which rapid production of novel therapeutics is needed. In plants, can be used as a gene delivery vector for transient expression. A potential barrier for plant-based production of human therapeutics is that different glycosylation patterns are found on plant and mammalian proteins. Since glycosylation can affect the efficacy, safety and stability of a therapeutic protein, methods to control glycan structures and distributions in plant-based systems would be beneficial. In these studies, we performed -mediated transient expression in glycoengineered plant cell suspension cultures. To reduce the presence of plant-specific glycans on the product, we generated and characterized cell suspension cultures from β-1,2-xylosyltransferase and α-1,3-fucosyltransferase knockdown . An anthrax decoy fusion protein was transiently produced in these glycoengineered plant cell suspension cultures through co-culture with genetically engineered . The mass ratio of to plant cells used was shown to impact recombinant protein expression levels. N-glycosylation analysis on the anthrax decoy fusion protein produced in glycoengineered showed a dramatic reduction in plant-specific N-glycans. Overall, the results presented here demonstrate the feasibility of a simple, rapid and scalable process for transient production of recombinant proteins without plant-specific glycans in a glycoengineered plant cell culture host.
瞬时重组蛋白生产是一种有前途的替代稳定转基因系统的方法，特别是在需要快速生产新型治疗药物的紧急情况下。在植物中，可以用作瞬时表达的基因传递载体。植物生产人类治疗药物的一个潜在障碍是植物和哺乳动物蛋白上存在不同的糖基化模式。由于糖基化会影响治疗蛋白的功效、安全性和稳定性，因此控制植物系统中聚糖结构和分布的方法将是有益的。在这些研究中，我们在糖基化工程植物细胞悬浮培养物中进行了介导的瞬时表达。为了减少产物中植物特异性聚糖的存在，我们生成并表征了β-1,2-木糖基转移酶和α-1,3-岩藻糖基转移酶敲低的细胞悬浮培养物。通过与遗传工程化的共培养，在这些糖基化工程植物细胞悬浮培养物中瞬时生产炭疽诱饵融合蛋白。用于与植物细胞共培养的与植物细胞的质量比被证明会影响重组蛋白的表达水平。在糖基化工程植物细胞中生产的炭疽诱饵融合蛋白的 N-糖基化分析显示，植物特异性 N-聚糖显著减少。总体而言，这里呈现的结果证明了在糖基化工程植物细胞培养物宿主中，使用简单、快速和可扩展的方法瞬时生产无植物特异性糖基的重组蛋白是可行的。
