Improving process robustness of cation exchange chromatography with cationic buffers for the reduction of aggregates

IF 1.4 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Protein expression and purification Pub Date : 2024-12-27 DOI:10.1016/j.pep.2024.106657
Shuo Wang, Shuaihua Wang, Dijing Shi, Ruomei Lv
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Abstract

Cation exchange chromatography (CEX) is commonly used to separate aggregates from monomers during the industrial manufacturing of recombinant proteins. However, the similar isoelectric point of aggregates and monomers makes the stepwise elution CEX an unstable process. In this study, the performance robustness of sodium chloride stepwise elution and cationic buffers (histidine and Bis-Tris) stepwise elution were compared through Monte Carlo simulation. While all trials achieved acceptable levels of monomer purity, sodium chloride stepwise elution exhibited significant fluctuations in yield and elution volume due to variations in elution pH and eluent concentration. In contrast, histidine or Bis-Tris stepwise elution resulted in more consistent yield and elution volume across a broad operating range. The findings indicate that the dissociation behavior of cationic buffers mitigates the impact of pH variation on ion-exchange equilibrium and thus enables a more robust CEX process.
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用阳离子缓冲液减少聚集体,提高阳离子交换色谱的过程稳健性。
在重组蛋白的工业生产过程中,阳离子交换色谱(CEX)通常用于分离聚合体和单体。然而,聚集体和单体的等电点相似,使得逐步洗脱CEX的过程不稳定。本研究通过蒙特卡罗模拟比较了氯化钠逐步洗脱和阳离子缓冲液(组氨酸和Bis-Tris)逐步洗脱的性能稳健性。虽然所有试验都达到了可接受的单体纯度水平,但由于洗脱pH值和洗脱液浓度的变化,氯化钠逐步洗脱的产率和洗脱量出现了显著波动。相比之下,组氨酸或Bis-Tris逐步洗脱在广泛的操作范围内产生更一致的产量和洗脱量。研究结果表明,阳离子缓冲液的解离行为减轻了pH变化对离子交换平衡的影响,从而使CEX过程更加稳健。
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来源期刊
Protein expression and purification
Protein expression and purification 生物-生化研究方法
CiteScore
3.70
自引率
6.20%
发文量
120
审稿时长
32 days
期刊介绍: Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.
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