海洋科学2025，Vol.49Issue(6)：86-97,12.DOI:10.11759/hykx20250430001
Ⅱ型鲨鱼源单域抗体EF73在大肠杆菌中的稳定表达
Robust expression of the Type Ⅱ shark-derived single domain antibody EF73 in Escherichia coli
摘要
Abstract
Sharks produce a heavy-chain antibody known as immunoglobulin new antigen receptor(IgNAR),which lacks light chains.Its variable region,the variable domain of the new antigen receptor(VNAR),can independently bind to antigens.Together with variable domain of heavy-chain antibodies(VHHs),VNARs are referred to as sin-gle-domain antibodies.Robust expression of shark recombinant VNARs is crucial for their characterization and application.However,their expression and yield often suffer from significant batch-to-batch variability,and reports on the recombinant VNAR expression remain limited.In this study,we devised a strategy for optimizing the ex-pression of EF73,a Type Ⅱ shark-derived VNAR.Results showed that inoculating the initial culture with multiple colonies rather than a single colony,along with expressing EF73 as a TrxA fusion,consistently facilitated high-level expression of soluble EF73 in both the periplasm and cytoplasm.This strategy provides a practical approach to overcoming the challenges associated with VNAR expression and may promote further research and application of shark-derived single-domain antibodies.关键词
鲨鱼源单域抗体/原核表达系统/蛋白重组表达Key words
variable domain of the new antigen receptor/prokaryotic expression system/recombinant protein expression分类
生物科学引用本文复制引用
丛思璇,邓鹏辉,徐明策,闻建晴,王淏,毕允晨..Ⅱ型鲨鱼源单域抗体EF73在大肠杆菌中的稳定表达[J].海洋科学,2025,49(6):86-97,12.基金项目
国家自然科学基金(42306144) （42306144）
青岛市关键技术攻关及产业化示范项目(25-1-1-gjgg-76-nsh)the National Natural Science Foundation of China,No.42306144 （25-1-1-gjgg-76-nsh）
Key Technology Research and Industrialization Demonstration Project of Qingdao,No.25-1-1-gjgg-76-nsh （）
