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Updated: Jun 27, 2025

Split-BioID — Proteomic Analysis of Context-specific Protein Complexes in Their Native Cellular Environment

Split-BioID — Proteomic Analysis of Context-specific Protein Complexes in Their Native Cellular Environment

Published on: April 20, 2018

使用新的分裂蛋白亲缘关系技术生产原生重组蛋白质

Robert Clifford¹, Susanna Lindman², Jie Zhu³

¹Purification Process Sciences, Process and Analytical Sciences, R&D Biopharmaceuticals, AstraZeneca LLC, One Medimmune Way, Gaithersburg, MD 20878, USA.

Journal of chromatography. A

|April 26, 2024

概括

这项研究介绍了CytivaTM ProteinSelectTM,这是一个新的分裂蛋白系统,用于高效的重组蛋白净化. 它可以在没有蛋白酶的情况下进行标签去除,并为大规模蛋白质生产提供强大的,可重复使用的亲和树脂.

科学领域:

生物技术是生物技术.
蛋白质化学蛋白质化学
亲密关系净化净化

背景情况:

亲和标签有助于重组蛋白净化,但由于生物干扰和免疫性,通常需要去除.
传统的标签去除需要昂贵的蛋白酶,这对成本有效的蛋白质生产构成了挑战.
分裂蛋白质系统提供了一个无蛋白质酶的替代方案,通过使蛋白质捕获后的自我切割.

研究的目的:

评估新型CytivaTM ProteinSelectTM 分割整体系统的应用和优势.
为了证明该系统对大规模蛋白质净化和树脂重复利用的能力.
使用该系统评估优化要求,以有效地表达蛋白质和恢复蛋白质.

主要方法:

使用CytivaTM ProteinSelectTM系统捕获SARS-CoV-2尖端蛋白受体结合域和双特异性T细胞参与者.
在多个循环中使用氧化进行了树脂再生和消毒的研究.
使用IL-1b和IFNAR-1 ECD进行了结合研究,以评估长期结合能力和可重复使用性.

主要成果:

成功捕获并扩大了目标蛋白质 (SARS-CoV-2尖端蛋白 RBD,双特异性T细胞参与者) 的生产10倍.
经证明强大的树脂结合能力 (10-20 g/L) 和在50次再生/消毒循环后保持能力.
在多次消毒循环后,确认保留了对目标蛋白的结合能力,突出了树脂的耐用性.

结论:

CytivaTM ProteinSelectTM系统是学术和工业蛋白质净化的一个可行和有利的工具.
该系统提供强大的蛋白质捕获,高效的标签去除,以及耐用,可重复使用的亲和树脂.
优化蛋白质N端序列和表达条件对于最大限度地提高切割效率和恢复至关重要.

关键词:

亲属性染色学是一种亲属性染色学.生物制药的发展.蛋白质净化蛋白质的净化

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