中国兽医科学2025，Vol.55Issue(9)：1166-1173,8.DOI:10.16656/j.issn.1673-4696.2025.0158
猪德尔塔冠状病毒重组N蛋白表达及单克隆抗体制备
Expression of porcine delta coronavirus recombinant N protein and generation of monoclonal antibody
摘要
Abstract
To prepare porcine deltacoronavirus(PDCoV)N protein and monoclonal antibodies(mAbs)against N protein,the PDCoV N gene was cloned into pGEX-6p-1 vector and transformed into BL21(DE3)competent cells.After induction and purification,recombinant N protein was obtained,about 43 kDa,and the size was as expected.Mice were immunized with the purified recombinant protein,and hybridoma cell lines producing anti-N protein mAbs were generated through cell fusion technology.Five stable hybridoma clones(2D8,2E2,6A1,9C10,and 5H10)exhibiting sustained antibody secretion were isolated.Immunological characterization using Western-blot and indirect immunofluorescence assay(IFA)confirmed the specific recognition of PDCoV N pro-tein by all five mAbs.Among them,2 strains(2D8,6A1)had a titer of ascites antibody as high as 1∶1 024 000.Neutralization test showed that both strains of monoclonal antibodies had a virus neutralization effect.This study achieved the successful prokaryotic expression of PDCoV N protein and established a panel of five mAbs with high specificity and neutralizing capacity.These findings provide critical tools for elucidating the functional role of PDCoV N protein and advancing diagnostic assay development.The generated reagents hold substantial potential for enhancing clinical diagnostics,epidemiological surveillance,and targeted disease control strategies for PDCoV infections.关键词
猪德尔塔冠状病毒/N蛋白/原核表达/单克隆抗体Key words
porcine deltacoronavirus/N protein/prokaryotic expression/monoclonal antibodies分类
农业科技引用本文复制引用
康欣桐,隋泽,赵梓桥,乌日尼乐,张昆丽,范瑞文,张建峰..猪德尔塔冠状病毒重组N蛋白表达及单克隆抗体制备[J].中国兽医科学,2025,55(9):1166-1173,8.基金项目
国家重点研发计划项目(2022YFD1800801-02) （2022YFD1800801-02）
广东省自然科学基金面上项目(2023A1515012153) （2023A1515012153）
山西省科技创新人才团队专项资助项目(202204051001022) （202204051001022）
猪禽种业全国重点实验室项目(2023QZ-NK05,GDNKY-ZQQZ-K07) （2023QZ-NK05,GDNKY-ZQQZ-K07）
科技创新战略专项资金(高水平农科院建设)-人才项目(R2023PY-QYB013) （高水平农科院建设）
