Valproic acid: A viable alternative to sodium butyrate for enhancing protein expression in mammalian cell cultures
Valproic acid: A viable alternative to sodium butyrate for enhancing protein expression in mammalian cell cultures
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DOI：
10.1002/bit.21882
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发表时间：
2008-09-01
影响因子：
3.8
通讯作者：
Wurm, Florian M.
中科院分区：
文献类型：
--
作者：
Backliwal, Gaurav;Hildinger, Markus;Wurm, Florian M.
研究了多种DNA甲基转移酶抑制剂(IDNMTs)和组蛋白脱乙酰酶抑制剂(IHDACs)对人胚胎肾293-EBNA(HEK293E)细胞瞬时基因表达的影响。将其在HEK293E细胞和中国仓鼠卵巢DG44(CHODG44)细胞中的作用进行了比较。根据iDNMTs和iHDACs不同的细胞活性和作用机制选择iDNMTs和iHDACs。对于所测试的每种抑制剂，确定了两种细胞系的最佳浓度，并利用重组单抗作为报告蛋白来评估每种化合物的效果。所有iHDAC在HEK293E和CHO-DG44中的瞬时抗体产量分别增加了至少4倍和1.5倍。相比之下，iDNMTs使抗体产量最多增加了-2倍。IDNMTs和iHDAC的配对组合对CHODG44的TGE有线性相加作用，而对HEK293E没有线性相加效应。在HEK293E细胞中加入丙戊酸(VPA)10天后，其体积产率和比产量分别达到200 mg/L和20pg/cell/d。由于VPA既是FDA批准的，又比丁酸钠(NaBut)便宜5倍，我们建议将其作为从哺乳动物细胞生产重组蛋白的广泛使用的促进剂的成本效益替代品。生物技术。比昂斯。2008年；101：182-189。(C)2008年威利期刊公司。
Various DNA methyl transferase inhibitors (iDNMTs) and histone deacetylase inhibitors (iHDACs) were screened for their ability to enhance transient gene expression (TGE) in Human Embryonic Kidney 293-EBNA (HEK293E) cells. The effects in HEK293E cells were compared to those in Chinese Hamster Ovary DG44 (CHODG44) cells. The iDNMTs and iHDACs were chosen based on their different cellular activities and mechanisms of action. For each inhibitor tested, the optimum concentration was determined for both cell lines, and these conditions were used to evaluate the effect of each compound using a recombinant monoclonal antibody as a reporter protein. All the iHDACs increased transient antibody yield at least 4-fold in HEK293E and at least 1.5-fold in CHO-DG44. By comparison, the iDNMTs increased antibody yields by a maximum of -2-fold. Pairwise combinations of iDNMTs and iHDACs had a linearly additive effect on TGE in CHODG44 but not in HEK293E. With valproic acid (VPA), volumetric and specific productivities of 200 mg/L and 20 pg/cell/day, respectively, were achieved in HEK293E cells with a 10-day process. As VPA is both FDA-approved and 5-fold less expensive than sodium butyrate (NaBut), we recommend it as a cost-effective alternative to this widely used enhancer of recombinant protein production from mammalian cells. Biotechnol. Bioeng. 2008;101: 182-189. (C) 2008 Wiley Periodicals, Inc.