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使用核糖体显示和Brevibacillus choshinensis分泌系统对抗体变异的评估

Monami Kihara¹, Rio Okuda², Anri Okada¹

¹Laboratory of Molecular Biotechnology, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.

Journal of bioscience and bioengineering

|March 22, 2025

概括

本研究提出了一种快速的方法,用于抗体工程,使用蛋白质合成,使用重组元素 (PURE) 核糖体显示和下一代测序 (NGS). 这种方法有效地识别高亲和度抗体变体,以改善治疗开发.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
免疫学免疫学免疫学

背景情况:

抗体工程需要有效的方法来增强抗体亲和力.
开发快速查策略对于优化基于抗体的治疗方法至关重要.

研究的目的:

建立一种用于高亲和抗体变异的高效丰富和分析的联合方法.
使用重组元素 (PURE) 核糖体显示器结合下一代测序 (NGS) 和Brevibacillus choshinensis分泌系统来利用蛋白质合成.

主要方法:

一个突变的单链片段变量 (scFab) 库是基于NZ-1抗体结构设计的.
在图书馆上进行了一轮PURE核糖体显示选择.
使用下一代测序 (NGS) 来分析所选变体的高丰富度和读数.

主要成果:

使用集成的PURE核糖体显示和NGS方法成功识别了高亲和度scFab候选者.
通过B. choshinensis分泌系统产生的顶级候选Fab表现出与野生类型相似的亲和力 (K_D = 1.6 × 10-9 M).

结论:

该研究表明,将PURE核糖体显示与NGS分析结合起来,以快速改善抗体亲和力的有效性.
布雷维细菌 (Brevibacillus choshinensis) 分泌系统是一个可行的平台,可以产生通过这种方法识别的高亲和度抗体变体.

关键词:

反-PA标签抗体的抗体.抗体工程是一种抗体工程.布雷维细菌 (Brevibacillus choshinensis) 是一种植物.没有细胞的蛋白质合成.核糖体显示器显示核糖体.

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