Laboratoire des Sciences du Génie Chimique, CNRS, 2 avenue de la Forêt-de-Haye, BP172, 54505, Vandoeuvre-lès-Nancy, France.
Cytotechnology. 2003 Jul;42(2):75-85. doi: 10.1023/B:CYTO.0000009816.65227.84.
The Baculovirus Expression Vector System has become widely used for the production of recombinant proteins for research and diagnostics. Serum-free culture media able to support high cell densities have been developed for the large scale culture of insect cells. While serum elimination aims at avoiding the risks associated with the introduction of an ill defined component of bovine origin, additives such as protein hydrolysates from animal sources are still used. An alternative could be the supplementation of culture media with protein hydrolysates derived from plants. In this study, we describe the replacement of lactalbumin hydrolysate with a laboratory produced hydrolysate of rapeseed proteins. Its effect on Sf9 cell growth kinetics, substrate consumption and by-product formation in low-serum or serum-free medium was evaluated. Cells were unable to grow in the presence of a rapeseed protein hydrolysate generated by PTN 3.0 Special((R)) enzyme and containing only 24% of peptides under 1 kDa in size. On the other hand, serum-free medium supplementation with a rapeseed protein hydrolysate obtained with Orientase 90N((R)) enzyme had a strong growth promoting effect, leading to a 60% increase in maximal cell density without affecting cell metabolism. This significant positive effect could be explained by the higher degree of hydrolysis of this digest, with 74% of peptides under 1 kDa in size.
杆状病毒表达载体系统已广泛用于生产用于研究和诊断的重组蛋白。已经开发出能够支持昆虫细胞大规模培养的无血清培养基。虽然消除血清旨在避免引入来源不明的牛成分带来的风险，但仍在使用来自动物来源的蛋白质水解物等添加剂。另一种选择可以是用植物来源的蛋白质水解物补充培养基。在这项研究中，我们描述了用实验室生产的油菜籽蛋白水解物替代乳白蛋白水解物。评估了它对 Sf9 细胞生长动力学、低血清或无血清培养基中基质消耗和副产物形成的影响。当存在由 PTN 3.0 Special((R))酶生成且仅含有 24%小于 1 kDa 大小的肽的油菜籽蛋白水解物时，细胞无法生长。另一方面，用 Orientase 90N((R))酶获得的油菜籽蛋白水解物补充无血清培养基具有很强的促生长作用，导致最大细胞密度增加 60%，而不影响细胞代谢。这种显著的积极影响可以通过这种消化物更高的水解程度来解释，其中 74%的肽小于 1 kDa。
