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Updated: May 5, 2026

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag

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Published on: January 16, 2012

使用卡帕选择和蛋白质L的多特异抗体的基于亲和的快速净化

Kalie Mix¹, Tingwan Sun¹, Brian Hall¹

¹Sanofi US Large Molecule Research, Cambridge, MA, USA.

|March 28, 2025

概括

这项研究引入了针对多种类型抗体 (msAbs) 的净化激活突变 (PEMs). 这些突变促进了与产品相关的杂质的去除,改善了治疗性抗体的产生.

科学领域:

生物技术是生物技术.
蛋白质工程是指蛋白质工程.
生物制药制造业生物制药制造业

背景情况:

多特异性抗体 (msAbs) 通过向多个生物分子提供治疗优势.
msAbs的生产面临着与产品相关的杂质的挑战,使净化复杂化,并可能降低产量.
现有的净化方法很难有效地去除特定的杂质,而不损害msAbs的完整性.

研究的目的:

开发一种多功能策略,用于在msAb净化过程中去除与产品相关的杂质.
设计msAbs的轻链,以调节它们与净化树脂的结合亲和力.
为两种特异性和三种特异性抗体建立基于亲和性的净化方案.

主要方法:

氨基酸突变被引入光链的恒定光链域和框架1.
改变的光链被测试了与卡帕选择 (KS) 和蛋白质L (Pro-L) 树脂的结合亲和力.
使用蛋白质A (Pro-A),KS和Pro-L树脂的多步亲和性净化过程被设计并应用于Y形双特异性和CODV三特异性抗体.

主要成果:

净化支持突变 (PEMs) 取消了与KS和Pro-L树脂的轻链结合,而不会影响抗原结合亲和力.
PEMs没有影响测试蛋白质的热稳定性或整体纯度.
一个基于亲和力的净化方案有效地消除了双特异性和三特异性抗体格式的轻链错配对物种.

结论:

通过选择性去除杂质,PEMs提供了一种强大的方法来增强msAbs的净化.
开发的基于亲和力的净化策略可以适应各种IgG类的msAb格式,包括具有Fc和Fab修改的格式.
这种方法适用于早期研究中的各种生产规模和高通量净化工作流程.

关键词:

双特异性抗体双特异性抗体在CODV中,CODV是CODV.具有多种特异性的抗体.蛋白质净化蛋白质净化三种特异性的抗体.

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