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在搅拌式反应器和气升式生物反应器中,利用 saUFL-AG-286 悬浮细胞系的无血清悬浮培养生产 Anticarsia gemmatalis 多角体病毒。

Production of the Anticarsia gemmatalis multiple nucleopolyhedrovirus in serum-free suspension cultures of the saUFL-AG-286 cell line in stirred reactor and airlift reactor.

机构信息

Laboratorio de Virología, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral, Santa Fe, Argentina.

出版信息

J Virol Methods. 2011 Dec;178(1-2):106-16. doi: 10.1016/j.jviromet.2011.08.024. Epub 2011 Aug 30.

Abstract

The velvetbean caterpillar, Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), is one of the main plagues for soybean crops. Velvetbean caterpillar larvae are susceptible to be infected by occlusion bodies of the baculovirus Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV), a biological insecticide. The insect cell line saUFL-AG-286 produces very high yields of occlusion bodies of AgMNPV in suspension cultures done in the low-cost serum-free medium UNL-10 in shake-flasks. However, its ability to adapt to conditions of industrial production in bioreactors was unknown. The aim of this study was to characterize the growth of saUFL-AG-286 cell cultures in UNL-10 medium, as well as its capability to replicate AgMNPV in two different bio-reactors at laboratory scale. The cell line was able to adapt to conditions that can be used at industrial scale, both in an airlift reactor and a stirred reactor, although the former was better than the last to support the cell growth. The infection with AgMNPV in the airlift reactor produced a high yield of occlusion bodies, with very low production of budded virus, the progeny used as inoculums. On the other hand, infection in the stirred reactor yielded high titers of budded virus. These results suggest that a feasible strategy for scaling-up the production of AgMNPV might involve the use of airlift reactors for the scaling-up of cell suspension cultures and the final production of occlusion bodies, while the scaling-up of the viral inoculums being carried out under conditions as those existing in stirred reactors.

摘要

绒泡桐尺蠖,Anticarsia gemmatalis Hübner(鳞翅目:夜蛾科),是大豆作物的主要病虫害之一。绒泡桐尺蠖幼虫易被杆状病毒 Anticarsia gemmatalis 多角体病毒(AgMNPV)的包涵体感染,AgMNPV 是一种生物杀虫剂。昆虫细胞系 saUFL-AG-286 在摇瓶中的无血清低成本培养基 UNL-10 中悬浮培养时,能够产生大量的 AgMNPV 包涵体。然而,其在工业生产规模的生物反应器中适应的能力是未知的。本研究的目的是表征 saUFL-AG-286 细胞在 UNL-10 培养基中的生长情况,以及其在实验室规模的两种不同生物反应器中复制 AgMNPV 的能力。该细胞系能够适应工业规模的条件,无论是在气升式反应器还是搅拌式反应器中,尽管前者比后者更有利于细胞生长。在气升式反应器中感染 AgMNPV 产生了大量的包涵体,而产生的芽生病毒产量非常低,芽生病毒可用作接种物。另一方面,在搅拌式反应器中感染产生了高滴度的芽生病毒。这些结果表明,扩大 AgMNPV 生产的可行策略可能涉及使用气升式反应器来扩大细胞悬浮培养物和最终生产包涵体,而接种物的扩大则在搅拌式反应器中存在的条件下进行。

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