Protein A chromatography resin lifetime-impact of feed composition.
Protein A chromatography resin lifetime-impact of feed composition.
复制标题
Protein A 色谱树脂寿命-进料成分的影响。
DOI：
10.1002/btpr.2608
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发表时间：
2018
影响因子：
2.9
通讯作者：
Pathak M
中科院分区：
文献类型：
--
作者：
Pathak M
Protein A 色谱过程中的吸附剂寿命不易预测或理解，这是生物制药制造商需要解决的一个关键挑战。本文重点关注进料成分对典型琼脂糖基 Protein A 树脂在 50 个循环寿命周期内性能的影响。使用三种不同的饲料原料进行循环研究，饲料成分的含量不同，包括蛋白酶、组蛋白、DNA 和非组蛋白。测量过程和质量属性的变化。尽管在所有情况下颗粒孔隙率均有所降低，但未发现 DBC 在不同条件下发生变化。使用荧光光谱和 LC-MS/MS 来确定污垢对观察到的容量损失的贡献和程度。三种饲料材料之间的残留蛋白 A 配体密度和污染物沉积（HCP、残留 mAb 和 DNA）各不相同。在含有高浓度 HCP 和组蛋白的饲料材料中循环的树脂被发现具有更大程度的容量损失。性能损失、容量损失或对产品质量的影响的模式根据进料的不同而变化。结果表明，进料成分可能与树脂老化的速率和模式相关，作为改进工艺理解的基础。 © 2018 美国化学工程师生物技术学会。编者，34：412–419，2018
Adsorbent lifetime during protein A chromatography is not readily predicted or understood, representing a key challenge to be addressed for biopharmaceutical manufacturers. This article focuses on the impact of feed composition on the performance of a typical agarose‐based protein A resin across a lifetime of 50 cycles. Cycling studies were performed using three different feed materials with varying levels of feed components including proteases, histones, DNA, and nonhistone proteins. Changes in the process and quality attributes were measured. The DBCs were not seen to vary between conditions although there was a reduction in particle porosity in all cases. Fluorescence spectroscopy and LC‐MS/MS were used to identify the contribution and extent of fouling to the observed capacity loss. Residual protein A ligand density and deposition of foulants (HCP, residual mAb, and DNA) varied between the three feed materials. Resins cycled in feed materials containing high concentrations of HCP and histones were seen to have greater extents of capacity loss. The mode of performance loss, capacity loss, or impact on product quality was seen to vary depending on the feed material. The results indicate that feed material composition may be correlated to the rate and mode of resin aging as a basis for improved process understanding. © 2018 American Institute of Chemical EngineersBiotechnol. Prog., 34:412–419, 2018