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在恒化器中培养黑腹果蝇(S2)。

Culturing Drosophila melanogaster (S2) in a chemostat.

作者信息

Vieira Paula Bruzadelle, da Costa Bruno Labate Vale, de Fatima Pires Augusto Elisabeth, Tonso Aldo

机构信息

Laboratório de Células Animais, Escola Politécnica--Universidade de São Paulo, P.O. Box 61.548, São Paulo, SP, CEP 05524-970, Brazil,

出版信息

Biotechnol Lett. 2015 Mar;37(3):533-8. doi: 10.1007/s10529-014-1717-9. Epub 2014 Nov 4.

Abstract

Insect cells are used for the expression of complex proteins in products such as vaccines and biopharmaceuticals. Physiology of a Drosophila melanogaster (lineage S2), transfected to stably express rabies virus glycoprotein (RVGP), was studied in batch culture and in a chemostat with serum-free medium. In batch mode, the system reached 3 × 10(7) cells mL(-1) with specific growth rate of 1.5 d(-1) with RVGP at 2.50 µg L(-1). When operated continuously, three well-defined steady states were achieved at dilution rates (D) of 0.8, 0.5 and 0.2 d(-1). The residual glucose and glutamine concentrations and the cell yields on glucose and glutamine decreased at lower D. High values of substrate consumption for maintenance may explain this variation in yields. The results indicated that glucose is not the limiting substrate of this process.

摘要

昆虫细胞用于在疫苗和生物制药等产品中表达复杂蛋白质。在分批培养和无血清培养基的恒化器中,研究了稳定表达狂犬病病毒糖蛋白(RVGP)的黑腹果蝇(品系S2)的生理学。在分批模式下,该系统达到3×10⁷个细胞·mL⁻¹,RVGP浓度为2.50 μg·L⁻¹时比生长速率为1.5 d⁻¹。连续运行时,在稀释率(D)为0.8、0.5和0.2 d⁻¹时实现了三个明确的稳态。在较低的D下,残余葡萄糖和谷氨酰胺浓度以及基于葡萄糖和谷氨酰胺的细胞产率降低。维持所需底物消耗的高值可能解释了产率的这种变化。结果表明葡萄糖不是该过程的限制底物。

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