南京医科大学学报（自然科学版）2025，Vol.45Issue(11)：1598-1607,10.DOI:10.7655/NYDXBNSN250094
广谱中和新型冠状病毒及其变体的纳米抗体制备及功能研究
Nanobody preparation and functional studies for broad-spectrum neutralization of SARS-CoV-2 and its variants
摘要
Abstract
Objective:To produce broad-spectrum nanobodies(Nbs)targeting severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)and its variants,and establish a strategy for producing Nbs broad-spectrum neutralizing the virus and its various variants.Methods:Alpacas were immunized with the receptor-binding domain(RBD)of the SARS-CoV-2 wild-type strain(WT-RBD)to construct a nanobody phage library,and the library was screened by biopanning and ELISA with the RBD of the Delta variant(Delta-RBD)to select Nbs.The selected Nbs were expressed and purified using the Pichia pastoris system,and their properties and functions characterized by ELISA,biolayer interferometry,pseudovirus neutralization experiments,and molecular modeling and docking.Results:Four Nbs were obtained,all exibiting high affinity for WT-RBD,while also effectively neutralizing Delta-RBD and the RBD of the Omicron variant(Omicron-RBD),with Nb45 effectively neutralizing the pseudovirus of(Omicron-RBD).We also found that the mechanism of these Nbs for neutralizing SARS-CoV-2 and its variants may be related to interfering with the conserved sites on these RBD.Conclusion:The four Nbs obtained have great potential for prevention and treatment of SARS-CoV-2 and its variants,and the strategy established in this study can also be widely used for the preparation of Nbs broad-spectrum neutralizing viruses and their various variants.关键词
SARS-CoV-2/变体/策略/中和性纳米抗体/假病毒Key words
SARS-CoV-2/variants/strategy/neutralizing nanobody/pseudovirus分类
基础医学引用本文复制引用
汪佰成,倪晶怡,刘馨月,龚雪,季晨博..广谱中和新型冠状病毒及其变体的纳米抗体制备及功能研究[J].南京医科大学学报（自然科学版）,2025,45(11):1598-1607,10.基金项目
江苏省"333高层次人才培养工程"项目 （）
南京市妇幼保健院"十四五"青年人才项目 （）
