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通过补充确定区域移植技术生成具有不同物理化学和生物特性的子单链变量片段

Ngoc Minh Nguyen¹, Kiichi Nakao¹, Ryo Kobayashi¹

¹Faculty of Molecular Chemistry and Engineering, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.

Journal of bioscience and bioengineering

|August 28, 2024

概括

这项研究引入了一种新的补充确定区域 (CDR) 接种方法,用于子单链可变片段 (scFvs). 这种精细的技术成功地保持了抗原结合亲和力,并允许在工程抗体中具有多种物理化学性质.

科学领域:

生物技术是生物技术.
免疫学免疫学免疫学
蛋白质工程是指蛋白质工程.

背景情况:

在子单链可变片段 (scFvs) 中,传统的补充决定区域 (CDR) 接种可以导致抗原结合亲和力降低.
在正规CDR之外的氨基酸残留物可能在子scFvs的抗原识别中发挥关键作用.

研究的目的:

开发一种改进的CDR移植技术,用于生成具有定制抗原识别和物理化学性质的子scFvs.
研究重新定义的CDR和框架区域 (FR) 对抗体结合亲和力和功能的影响.

主要方法:

通过扩展CDRH1和CDRH2中的氨基酸残留来重新定义CDR和FR,基于子scFvs的序列对齐.
产生和表征CDR移植的scFvs与常见的CDR和各种FRs.
使用三明治ELISA和蛋白质L结合试验评估抗原结合活性.

主要成果:

与原始克隆相比,重新定义的CDR/FR策略成功地保持了抗原结合亲和力.
抗原结合活动在固体支上有所不同,这表明FRs影响了scFv的物理化学特性.
CDR移植的scFvs保留了蛋白质L结合能力,使其能够净化并显示出更好的溶解性.

结论:

开发的CDR移植方法是有效的创造子scFvs与保存的抗原特异性和多样化的物理化学特征.
这项技术为产生各种应用的定制抗体提供了一个有前途的方法.
该研究强调了在抗体工程中考虑非正规区域的重要性.

关键词:

确定互补性的地区.补充-确定区域-接种.框架区域是指框架区域的区域.物理化学特性物理化学特性蛋白质L-结合的蛋白质子单链变量碎片 scFv 溶解度溶解度在

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