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通过添加蛋白质稳定剂以及从锥形瓶转移至2升生物反应器,烟草培养悬浮细胞中14D9催化抗体的表达量增加了。

The expression of the 14D9 catalytic antibody in suspended cells of Nicotiana tabacum cultures increased by the addition of protein stabilizers and by transference from Erlenmeyer flasks to a 2-L bioreactor.

作者信息

Marconi Patricia L, Alvarez María Alejandra

机构信息

Cátedra de Farmacobotánica y Farmacognosia, Carreras de Farmacia y Bioquímica, Facultad de Ciencias de la Salud, Universidad Maimónides, Ciudad de Buenos Aires, Argentina.

出版信息

Biotechnol Prog. 2014 Sep-Oct;30(5):1185-9. doi: 10.1002/btpr.1940. Epub 2014 Jul 1.

Abstract

The effect of two protein stabilizers (polyvinylpyrrolidone [PVP] and gelatine) on growth and 14D9 yield of Nicotiana tabacum cell suspension cultures (Ab-KDEL and sec-Ab) was analyzed. The addition of PVP at a concentration of 1.0 g L(-1) produced the highest total 14D9 yield (biomass + culture medium) in the Ab-KDEL line (4.82% total soluble protein [TSP]). With the addition of gelatine, the highest total 14D9 yield (2.48% TSP) was attained in the Ab-KDEL line at 5.0 g L(-1) gelatine. When the Ab-KDEL suspended cells were cultured in a 2-L bioreactor, the highest 14D9 yield was 8.1% TSP at a 5% w/v inoculum size, which was the best 14D9 yield so far obtained in the platforms tested (E. coli, N. tabacum leaves and seeds, N. tabacum hairy roots, and cell suspension cultures).

摘要

分析了两种蛋白质稳定剂(聚乙烯吡咯烷酮[PVP]和明胶)对烟草细胞悬浮培养物(Ab-KDEL和sec-Ab)生长和14D9产量的影响。在Ab-KDEL系中,添加浓度为1.0 g L(-1)的PVP时,14D9的总产量(生物量+培养基)最高(占总可溶性蛋白[TSP]的4.82%)。添加明胶时,在明胶浓度为5.0 g L(-1)的Ab-KDEL系中,14D9的总产量最高(占TSP的2.48%)。当Ab-KDEL悬浮细胞在2-L生物反应器中培养时,接种量为5% w/v时,14D9的最高产量为8.1% TSP,这是目前在测试平台(大肠杆菌、烟草叶片和种子、烟草毛状根和细胞悬浮培养物)中获得的最佳14D9产量。

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