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A Nanobody of PEDV S1 Protein: Screening and Expression in Escherichia coli

大肠杆菌 蛋白质表达 生物 病毒学 微生物学 分子生物学 化学 遗传学 基因
作者
Zhipeng Hao,Xufeng Dong,Qian Zhang,Zhihua Qin
出处
期刊:Biomolecules [MDPI AG]
卷期号:14 (9): 1116-1116
标识
DOI:10.3390/biom14091116
摘要

Porcine epidemic diarrhea virus (PEDV) has caused significant economic losses to the pig farming industry in various countries for a long time. Currently, there are no highly effective preventive or control measures available. Research into the pathogenic mechanism of PEDV has shown that it primarily causes infection by binding the S protein to the CD13 (APN) receptor on the membrane of porcine intestinal epithelial cells. The S1 region contains three neutralization epitopes and multiple receptor-binding domains, which are closely related to viral antigenicity and ad-sorption invasion. Nanobodies are a type of single-domain antibody that have been discovered in recent years. They can be expressed on a large scale through prokaryotic expression systems, which makes them cost-effective, stable, and less immunogenic. This study used a phage display library of nanobodies against the PEDV S1 protein. After three rounds of selection and enrichment, the DNA sequence of the highly specific nanobody S1Nb1 was successfully obtained. To obtain soluble nanobody S1Nb1, its DNA sequence was inserted into the vector Pcold and a solubility-enhancing SUMO tag was added. The resulting recombinant vector, Pcold-SUMO-S1Nb1, was then transformed into
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