Abstract

Yeast surface display is a powerful technology used to isolate and engineer proteins to improve their activity, specificity, and stability. In this method, gene expression is regulated by promoters, and secretion efficiency is affected by secretion signals. Furthermore, both the accessibility and activity of the displayed proteins are affected by the length of anchor proteins. The ideal promoter, secretion signal, and anchor protein combination depend on the proteins of interest. In this study, we optimized a yeast surface display suitable for nanobody evaluation. We designed five display systems that used different combinations of promoters, secretion signals, and anchor proteins. Anti-hen egg-white lysozyme nanobody was used as the model nanobody. The amount of nanobodies displayed on yeast cells, the number of antigens bound to the displayed nanobodies, and the display efficiency were quantified. Overall, we improved the yeast display system for nanobody engineering and proposed its optimization.

中文翻译：

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用于筛选功能性纳米抗体的酵母表面展示系统的评估

摘要

酵母表面展示技术是一项强大的技术，可用于分离和改造蛋白质以提高其活性，特异性和稳定性。在该方法中，基因表达由启动子调节，并且分泌效率受分泌信号影响。此外，展示蛋白的可及性和活性均受锚蛋白长度的影响。理想的启动子，分泌信号和锚蛋白组合取决于目标蛋白。在这项研究中，我们优化了适用于纳米抗体评估的酵母表面展示。我们设计了五个显示系统，它们使用启动子，分泌信号和锚蛋白的不同组合。抗鸡卵清溶菌酶纳米抗体被用作模型纳米抗体。酵母细胞上展示的纳米抗体的数量，与展示的纳米抗体结合的抗原数量和展示效率被定量。总体而言，我们改进了用于纳米抗体工程的酵母展示系统，并提出了其优化方案。