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Updated: Jun 10, 2025

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具有高度选择性的分裂蛋白质方法,可有效地从哺乳动物细胞培养液中分离和净化重组治疗蛋白质

Sai Vivek Prabhala¹, Brian Marshall², Jackelyn Galiardi³

¹William G. Lowrie Department of Chemical and Biomolecular Engineering, Ohio State University, Columbus, OH 43210, United States.

Journal of chromatography. A

|October 15, 2024

概括

一个新的自我清除的亲和度标签,iCapTagTM,为净化重组蛋白提供了一个通用平台. 该系统可在单个步骤中实现高效,无标签的蛋白质净化,简化了临床应用的制造.

科学领域:

生物技术是生物技术.
蛋白质净化蛋白质净化
亲和染色学是一种亲和染色学.

背景情况:

治疗性蛋白质需要高纯度用于临床使用,但当前的净化方法,如单克隆抗体的蛋白A,并不普遍适用.
现有的多列色谱过程是复杂的,需要广泛的优化,并降低最终产品的产量.
对于各种重组蛋白质净化,需要一个广泛适用的亲和平台.

研究的目的:

介绍和演示新的自我清除iCapTagTM作为复合蛋白质的通用捕获和净化平台.
展示iCapTagTM在生产适合临床使用的无标签治疗蛋白质方面的效率和适用性.
为了验证不同重组蛋白类型的iCapTagTM性能.

主要方法:

开发了iCapTagTM系统,该系统可以捕获,缩和净化蛋白质.
使用简单的缓冲器pH转移来删除标签,不需要额外的试剂或辅助因子.
涉及干扰素α2b,ML39单链可变片段 (scFv) 和Expi293细胞表达的SARS-CoV-2尖端蛋白受体结合域 (RBD) 净化的案例研究.

主要成果:

该iCapTagTM系统使无标签蛋白质的高效净化 (>99%纯度) 在单个步骤中成为可能.
证明了宿主细胞DNA,标记前体和分子量变异的高清除率.
成功净化各种蛋白质,包括干扰素α2b,scFv和RBD,所有这些都产生了无标签的最终产品.

结论:

该iCapTagTM功能作为捕获,缩和净化重组蛋白质的多功能和高效平台.
标签的自我清除性质,由pH值变化引发,简化了该过程,并产生适合治疗应用的无标签蛋白质.
iCapTagTM为重组蛋白制造提供了一个有前途的通用解决方案,解决了当前净化策略的局限性.

关键词:

生物分离生物分离染色体学染色体学是一种染色体学.下游加工是下游加工.在英特因的基础上.蛋白质净化蛋白质的净化自切割的亲和度标签自切割亲和度标签

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