Reduction of chromatin heteroaggregates by acid precipitation of mammalian cell culture and ramification in protein A chromatography for recombinant immunoglobulin G purification
Reduction of chromatin heteroaggregates by acid precipitation of mammalian cell culture and ramification in protein A chromatography for recombinant immunoglobulin G purification
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通过哺乳动物细胞培养物的酸沉淀和蛋白 A 层析中的分支来减少染色质杂聚体，用于重组免疫球蛋白 G 纯化
DOI：
10.1016/j.bej.2018.12.012
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发表时间：
2019-03-15
影响因子：
3.9
通讯作者：
Nian, Rui
中科院分区：
文献类型：
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作者：
Liu, Wenshuai;Zhou, Dongyan;Nian, Rui
酸沉淀已被证明是免疫球蛋白G（IgG）纯化过程中细胞培养上清液（CCS）预处理的一种澄清方法。在这项研究中，IgG，非组蛋白宿主细胞蛋白（n-hHCP），组蛋白，和DNA的反应，在不同的pH值的盐（NaCl）的存在下，在不同的浓度进行了系统的研究。IgG的丢失与染色质异聚集体的沉淀密切相关，其中DNA/组蛋白随缓冲液环境的酸化而沿着显著降低。此外，pH中和后DNA显示出完全恢复，而低pH处理诱导的组蛋白沉淀是永久性的，并且不能随着pH升高而恢复。分析性尺寸排阻色谱（SEC）图谱进一步表明染色质异聚集体的聚集和减少沿着较低的pH。蛋白A色谱受益于染色质异聚集体的减少，并显示出杂质去除以及IgG回收的显著增强。本研究首次将CCS酸化与染色质异聚集体去除联系起来，并说明了其对后续纯化过程的影响。
Acid precipitation has been demonstrated as a clarification method for cell culture supernatant (CCS) pre-treatment during Immunoglobulin G (IgG) purification. In this study, the responses of IgG, non-histone host cell protein (n-hHCP), histone, and DNA to different pHs in the presence of salt (NaCl) at various concentrations were systematically studied. IgG loss was closely associated with the precipitation of chromatin heteroaggregates, in which DNA/histone showed significant decreases along with the acidification of buffer environment. In addition, DNA showed full recovery after pH neutralization, while the low pH treatment induced histone precipitation was permanent and could not be reverted upon pH increase. Analytical size exclusion chromatography (SEC) profiles further indicated the agglomeration and reduction of chromatin heteroaggregates along with lower pH. Protein A chromatography benefited from the reduction of chromatin heteroaggregates and showed significant enhancement of the removal of impurities as well as for IgG recovery. For the first time, this study correlated CCS acidification with chromatin heteroaggregate removal and illustrated its ramification for the subsequent purification process.