Center for Agricultural Biotechnology, University of Maryland, College Park, Maryland 20742.
Biotechnol Bioeng. 1993 Jun 20;42(2):240-6. doi: 10.1002/bit.260420212.
Insect cell culture and the baculovirus vector expression system have emerged to be a promising production technique for heterologous proteins. In this article, expression characteristics for membrane-bound epoxide hydrolase are examined. A generic process is presented whereby cells are grown in serum-free media supplemented with serum and then resuspended in serum-free media to simplify purification after infection. The infected cells retain significant metabolic activity during the postinfection stage. Thus, maintaining nutrient supply during the postinfection period is critical, and a low stirring rate will result in oxygen depletion and shift the metabolism of the infected cells toward lactate production which then lowers product yield. This is the first report indicating that glucose is supplied from sucrose decomposition and then metabolized for viral DNA and recombinant protein production in recombinant baculovirus insect expression system.
昆虫细胞培养和杆状病毒载体表达系统已经成为一种很有前途的异源蛋白生产技术。本文研究了膜结合环氧化物水解酶的表达特性。提出了一种通用的工艺，即在无血清培养基中培养细胞，并补充血清，然后在感染后将细胞重悬于无血清培养基中，以简化感染后的纯化过程。感染后的细胞在感染后阶段仍保持显著的代谢活性。因此，在感染后阶段维持营养供应是至关重要的，低搅拌速率会导致氧气耗尽，并使感染细胞的代谢转向乳酸生产，从而降低产物产量。这是首次表明在重组杆状病毒昆虫表达系统中，葡萄糖是由蔗糖分解提供的，然后用于病毒 DNA 和重组蛋白的生产。
