Pathak Mili, Rathore Anurag S, Lintern Katherine, Bracewell Daniel G
Department of Chemical Engineering, Indian Institute of Technology, New Delhi, India.
Department of Biochemical Engineering, University College London, Gower, Street, UK London.
Biotechnol Prog. 2018 Mar;34(2):412-419. doi: 10.1002/btpr.2608. Epub 2018 Jan 26.
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 Engineers Biotechnol. Prog., 34:412-419, 2018.
在蛋白A层析过程中，吸附剂的使用寿命难以预测或理解，这是生物制药制造商需要解决的一个关键挑战。本文重点研究了在50个循环周期内，进料组成对典型琼脂糖基蛋白A树脂性能的影响。使用三种不同的进料材料进行循环研究，这些进料材料中进料成分的含量各不相同，包括蛋白酶、组蛋白、DNA和非组蛋白。测量了工艺和质量属性的变化。尽管在所有情况下颗粒孔隙率都有所降低，但未观察到动态结合容量（DBC）在不同条件之间有所变化。使用荧光光谱和液相色谱-串联质谱（LC-MS/MS）来确定污垢对观察到的容量损失的贡献和程度。三种进料材料之间的残留蛋白A配体密度和污垢（宿主细胞蛋白、残留单克隆抗体和DNA）的沉积情况各不相同。在含有高浓度宿主细胞蛋白和组蛋白的进料材料中循环的树脂，其容量损失程度更大。观察到性能损失的模式、容量损失或对产品质量的影响因进料材料而异。结果表明，进料材料组成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美国化学工程师学会生物技术进展，34:412 - 419，2018。
