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首页> 外文期刊>Bioprocess and Biosystems Engineering >Precipitation of complex antibody solutions: influence of contaminant composition and cell culture medium on the precipitation behavior
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Precipitation of complex antibody solutions: influence of contaminant composition and cell culture medium on the precipitation behavior

机译：复杂抗体溶液的沉淀：污染物组成和细胞培养基对沉淀行为的影响

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摘要

Preparative protein precipitation is known as a cost-efficient and easy-to-use alternative to chromatographic purification steps. This said, at the moment, there is no process for monoclonal antibodies (mAb) on the market, although especially polyethylene glycol-induced precipitation has shown great potential. One reason might be the highly complex behavior of each component of a crude feedstock during the precipitation process. For different investigated mAbs, significant variations in the host cell protein (HCP) reduction are observed. In contrast to the precipitation behavior of single components, the interactions and interplay in a complex feedstock are not fully understood yet. This work discusses the influence of contaminants on the precipitation behavior of two different mAbs, an IgG1, and an IgG2. By spiking the mAbs with mock solution, a complex feedstock could successfully be mimicked. Spiking contaminants influenced the yield and purity of the mAbs after the precipitation step, compared to the precipitation behavior of the single components. The mixture showed a decrease in the contaminant and mAb solubility. By re-buffering the mock solution prior to spiking, special salts, small molecules like amino acids, vitamins, or sugars could be depleted while larger ones like HCP or DNA were still present. Therefore, it was possible to distinguish the influence of small molecules and larger ones. Hence, mAb-macromolecular interaction could be identified as a possible reason for the observed higher precipitation propensity, while small molecules of the cell culture medium were identified as solubilisation factors during the precipitation process.
机译：制备蛋白沉淀是色谱纯化步骤的一种经济高效且易于使用的替代方法。这就是说，尽管特别是聚乙二醇诱导的沉淀显示出巨大的潜力，但目前市场上还没有单克隆抗体（mAb）的生产方法。一个原因可能是在沉淀过程中，原油原料中每种成分的行为非常复杂。对于研究的不同mAb，观察到宿主细胞蛋白（HCP）还原的显着变化。与单一组分的沉淀行为相反，复杂原料中的相互作用和相互作用尚不完全清楚。这项工作讨论了污染物对两种不同mAb（IgG1和IgG2）的沉淀行为的影响。通过用模拟溶液加标单克隆抗体，可以成功地模拟复杂的原料。与单个组分的沉淀行为相比，掺入杂质会影响沉淀步骤之后mAb的产率和纯度。该混合物显示出污染物和mAb溶解度的降低。通过在加标之前重新缓冲模拟溶液，可以消耗特殊的盐，氨基酸，维生素或糖等小分子，而仍存在HCP或DNA等大分子。因此，可以区分小分子和大分子的影响。因此，mAb-大分子相互作用可以被确定为观察到更高的沉淀倾向的可能原因，而细胞培养基的小分子被确定为沉淀过程中的增溶因子。

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