Betenbaugh M J, Balog L, Lee P S
Department of Chemical Engineering, Johns Hopkins University, Baltimore, Maryland 21218.
Biotechnol Prog. 1991 Sep-Oct;7(5):462-7. doi: 10.1021/bp00011a012.
An experimental study was undertaken to evaluate alternative insect cell lines to Sf9 [from Spodoptera frugiperda (fall armyworm)] for the production of recombinant proteins. Insect cell lines from two different organisms were considered: IPLB-LdEIta (LdEIta) from Lymantria dispar (gypsy moth) and IPLB-HvT1 (HvT1) from Heliothis virescens (tobacco budworm). Both LdEIta and HvT1 produced higher total activity levels of recombinant beta-galactosidase in monolayer culture than Sf9 after infection with the Autographa californica nuclear polyhedrosis virus (AcMNPV). However, only LdEIta generated a product yield (activity per milligram of total protein) which exceeded that of Sf9 (by 25%), so its growth and production characteristics were investigated in depth. LdEIta generated production levels and yields of a recombinant rotaviral protein, VP4, which exceeded those of Sf9 by 84 and 38%, respectively. In suspension culture, the LdEIta cells grew as aggregates with a doubling time several hours longer than Sf9, but the recombinant product yields of LdEIta were still higher than Sf9 by 38% in this culture environment. beta-Galactosidase expression rates and cell death rates suggested that the difference in productivity between the two hosts was due to the ability of LdEIta to survive the baculovirus infection and produce recombinant proteins longer than Sf9. The presence of LdEIta aggregates in suspension culture may be used as a method to separate live cells from dead cells, labile product, and spent medium in recombinant protein production processes.
进行了一项实验研究，以评估用于生产重组蛋白的替代昆虫细胞系，替代 Sf9 [来自草地贪夜蛾（秋粘虫）]。研究了来自两种不同生物体的昆虫细胞系：来自舞毒蛾的 IPLB-LdEIta（LdEIta）和来自烟草天蛾的 IPLB-HvT1（HvT1）。在用苜蓿银纹夜蛾核型多角体病毒（AcMNPV）感染后，LdEIta 和 HvT1 在单层培养中产生的重组β-半乳糖苷酶的总活性水平均高于 Sf9。然而，只有 LdEIta 的产物产量（每毫克总蛋白的活性）超过了 Sf9（高出 25%），因此对其生长和生产特性进行了深入研究。LdEIta 产生的重组轮状病毒蛋白 VP4 的产量和水平分别比 Sf9 高出 84%和 38%。在悬浮培养中，LdEIta 细胞聚集成团生长，其倍增时间比 Sf9 长几个小时，但在这种培养环境下，LdEIta 的重组产物产量仍比 Sf9 高 38%。β-半乳糖苷酶表达率和细胞死亡率表明，两种宿主之间生产力的差异是由于 LdEIta 能够在杆状病毒感染中存活并比 Sf9 更长时间地产生重组蛋白。悬浮培养中 LdEIta 聚集体的存在可作为一种方法，在重组蛋白生产过程中将活细胞与死细胞、不稳定产物和用过的培养基分离。
