In silico optimization of a challenging bispecific antibody chromatography step.

IF 2.5 3区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Zsófia Bencze, Tobias Hahn, Henri Kornmann, Pia Graf, Tatjana Trunzer
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引用次数: 0

Abstract

Mechanistic modeling of chromatographic steps is an effective tool in biopharma process development that enhances process understanding and accelerates optimization efforts and subsequent risk assessment. A relatively new model for ion exchange chromatography is the colloidal particle adsorption (CPA) formalism, which promises improved separation of material and molecule-specific parameters. This case study demonstrates a straightforward CPA modeling workflow to describe an ion exchange chromatography polishing step of a knobs-into-holes construct bispecific antibody molecule. An adapted Yamamoto method was used to calculate charge and equilibrium parameters at three pH values. The remaining model parameters, binding kinetics, and effective mass transfer coefficients were determined via inverse fitting. The model was created from six experiments in total, tested on model parameter uncertainty, and evaluated on its power to predict changes in the biomolecule's retention behavior when variations in elution salt concentration occur. Finally, a three-step-gradient experiment was optimized, separating the desired bispecific antibody from its low and high molecular weight impurities, achieving a monomer yield of 68% and purity of 96%. Testing the model against a different load composition demonstrated its ability to extrapolate. An in silico one-factor-at-time and two-parameter screening of the optimized method identified the salt concentration to elute weaker binding impurities as a critical process attribute, while deviations in the buffer pH had a minor influence.

一个具有挑战性的双特异性抗体色谱步骤的硅优化。
色谱步骤的机制建模是生物制药工艺开发的有效工具,可以增强对工艺的理解,加快优化工作和后续风险评估。一种相对较新的离子交换色谱模型是胶体颗粒吸附(CPA)形式,它有望改善物质和分子特异性参数的分离。本案例研究演示了一个简单的CPA建模工作流程来描述一个离子交换色谱抛光步骤的旋钮入孔构建双特异性抗体分子。采用改进的山本方法计算了三种pH值下的电荷和平衡参数。剩余的模型参数、结合动力学和有效传质系数通过逆拟合确定。该模型由总共6个实验创建,测试了模型参数的不确定性,并评估了其预测洗脱盐浓度变化时生物分子保留行为变化的能力。最后,优化三步梯度实验,将所需双特异性抗体从低分子量和高分子量杂质中分离出来,单体产率为68%,纯度为96%。对不同负载组成的模型进行测试,证明了它的推断能力。对优化方法进行了单因素单参数筛选,确定了洗脱较弱结合杂质的盐浓度是关键工艺属性,而缓冲液pH的偏差影响较小。
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来源期刊
Biotechnology Progress
Biotechnology Progress 工程技术-生物工程与应用微生物
CiteScore
6.50
自引率
3.40%
发文量
83
审稿时长
4 months
期刊介绍: Biotechnology Progress , an official, bimonthly publication of the American Institute of Chemical Engineers and its technological community, the Society for Biological Engineering, features peer-reviewed research articles, reviews, and descriptions of emerging techniques for the development and design of new processes, products, and devices for the biotechnology, biopharmaceutical and bioprocess industries. Widespread interest includes application of biological and engineering principles in fields such as applied cellular physiology and metabolic engineering, biocatalysis and bioreactor design, bioseparations and downstream processing, cell culture and tissue engineering, biosensors and process control, bioinformatics and systems biology, biomaterials and artificial organs, stem cell biology and genetics, and plant biology and food science. Manuscripts concerning the design of related processes, products, or devices are also encouraged. Four types of manuscripts are printed in the Journal: Research Papers, Topical or Review Papers, Letters to the Editor, and R & D Notes.
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