阴离子交换色谱纯化过程中小鼠微小病毒洗脱行为的机理模型。

IF 2.5 3区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology Progress Pub Date : 2024-10-23 DOI:10.1002/btpr.3516
Ryunosuke Kitamura, Lena Enghauser, Riku Miyamoto, Takahiro Ichikawa, Takaki Aiso, Yumiko Masuda, Daisuke Kajihara, Hirofumi Kakihara, Koichi Nonaka
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引用次数: 0

摘要

本研究旨在提出一种方法,用于建立流动阴离子交换(AEX)色谱法清除小鼠细小病毒(MVM)病毒的机理模型。蛋白质表面分析被用来研究重组生物治疗药物与MVM之间分子相互作用的可能性。在不含蛋白质产物的 Tris 缓冲液中添加 MVM,并使用定量聚合酶链反应(qPCR)对汇集馏分进行定量分析。GoSilico™ 色谱建模软件用于开发 MVM 物种的机理模型。为评估所建模型的视觉拟合度,各馏分中完整 MVM 病毒和未包被囊壳的模拟量与测量值之间的 R2 分别为 0.880 和 0.948。生成了负载样品中对数还原值(LRV)与 pH 值和电导率的响应面图,以显示实现良好 LRV 的合适负载样品条件的范围。为了评估所开发的 MVM 洗脱模型在重组生物治疗药物中的适用性,使用模型单克隆抗体的负载样品进行了两次 AEX 色谱纯化演示。所开发的模型准确地模拟了两次运行的洗脱步骤中 MVM 物种的峰值。此外,为了评估病毒与目标蛋白之间的分子相互作用是否会显著影响 MVM 的洗脱行为,还利用表面分析法对抗体表面的疏水性/亲水性进行了评估。
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Mechanistic model of minute virus of mice elution behavior in anion exchange chromatography purification.

This study aimed to propose a methodology for developing a mechanistic model for viral clearance of the minute virus of mice (MVM) on flow-through anion exchange (AEX) chromatography. Protein surface analysis was applied to investigate the possibility of molecular interaction between the recombinant biotherapeutic and MVM. The protein product-free Tris buffers were spiked with MVM, and the MVM elution profile from AEX chromatography was quantitatively analyzed using quantitative polymerase chain reaction (qPCR) for pooled fractions. GoSilico™ Chromatography Modeling Software was applied to develop the mechanistic models for MVM species. For evaluating the visual fit of the developed model, the R2 of intact MVM virions and uncoated capsids between the simulated and measured amount in each fraction are 0.880 and 0.948, respectively. Response surface plots of logarithmic reduction values (LRV) against pH and conductivity in loaded sample were generated to show the range for suitable loaded sample conditions for achieving a good LRV. To evaluate the applicability of the developed MVM elution model to a recombinant biotherapeutic, two demonstrations of AEX chromatography purification were performed with a loaded sample of a model monoclonal antibody. The peaks of the MVM species in the elution step of both runs were accurately simulated by the developed model. In addition, to assess the possibility of molecular interaction between the virus and the target protein significantly affecting the MVM elution behavior, the antibody's surface was evaluated in terms of hydrophobicity/hydrophilicity using surface analysis.

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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.
期刊最新文献
Non-thermal plasma decontamination of microbes: a state of the art. Mechanistic model of minute virus of mice elution behavior in anion exchange chromatography purification. Comparing in silico flowsheet optimization strategies in biopharmaceutical downstream processes. General strategies for IgG-like bispecific antibody purification. Issue Information
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