利用核糖体跳跃系统和耐热酵母优化酵母展示策略，以提高 scFv 抗体的筛选效率。

An optimized yeast display strategy for efficient scFv antibody selection using ribosomal skipping system and thermo resistant yeast.

机构信息

College of Life Sciences, Hebei Agricultural University, Baoding, 071001, Hebei, China.

Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, Shanghai, 201210, China.

出版信息

Biotechnol Lett. 2019 Sep;41(8-9):1067-1076. doi: 10.1007/s10529-019-02710-5. Epub 2019 Jul 12.

DOI:10.1007/s10529-019-02710-5

PMID:31300936

Abstract

OBJECTIVES

Establish a method to restrict unexpected fragments including stop codons in scFv library and generate a thermo resistant strain for screening of thermal stable scFv sequences.

RESULTS

Here, we have constructed a T2A-Leu2 system for selection of yeast surface display libraries that blocks amplification of "stop codon" plasmids within the library, thereby increasing the quality of the library and efficiency of the selection screen. Also, we generated a temperature-resistant yeast strain, TR1, and validated its combined use with T2A-Leu2 for efficient screening. Thus, we developed a general approach for a fast and efficient screening of scFv libraries using a ribosomal skipping system and thermo-resistant yeast.

CONCLUSIONS

The method highlights the utility of the T2A-Leu2-based ribosomal skipping strategy for increasing the quality of the input library for selection, along with an optimized selection protocol based on thermo-resistant yeast cells.

摘要

目的

建立一种方法来限制 scFv 文库中包含终止密码子的意外片段，并生成耐热菌株，以筛选热稳定 scFv 序列。

结果

在这里，我们构建了一个 T2A-Leu2 系统，用于选择酵母表面展示文库，该系统可以阻止文库中“终止密码子”质粒的扩增，从而提高文库的质量和选择筛选的效率。此外，我们还生成了一个耐热酵母菌株 TR1，并验证了其与 T2A-Leu2 的联合使用，以实现高效筛选。因此，我们开发了一种使用核糖体跳跃系统和耐热酵母快速高效筛选 scFv 文库的通用方法。