中国兽医科学2024，Vol.54Issue(9)：1209-1215,7.DOI:10.16656/j.issn.1673-4696.2024.0151
环形泰勒虫Tams1单个B细胞抗体的制备及鉴定
Preparation and identification of single B cell antibody against Tams1 of Theileria annulata
摘要
Abstract
The purpose of this study was to prepare specific monoclonal antibodies against Tams1 protein of Theileria annulata by single B cell antibody technology and to identify its reactivity.The pET-30a-Tams1 recombinant plasmid was constructed,and the protein was expressed and purified.The pu-rified Tams1 protein was used to immunize healthy cattle.After the fourth immunization,B lymphocytes secreting anti-Tams1-specific monoclonal antibodies(IgM-/CD21+/antigen+B lymphocytes)were obtained from peripheral blood mononuclear cells by flow sorting.After single cell lysis and reverse tran-scription,the light/heavy chain variable region sequence of IgG natural pairing was obtained by nested PCR amplification and cloned into the eukaryotic expression vector containing the constant re-gion of IgG light/heavy chain.After co-transfection of HEK-293 T cells with paired plasmids,the cell culture supernatant was collected.Western-blot results indicated that all the eight antibodies could bind to Tams1 protein.At the same time,IFA results showed that the eight antibodies could specifically bind to the merozoite surface antigen Tams1.Above results suggested that 8 strains of Tams1 specific monoclonal antibodies were successfully obtained in this study,which laid a foundation for the subse-quent function study of the protein and the establishment of a single B cell antibody preparation tech-nology platform.关键词
环形泰勒虫/Tams1/单个B细胞抗体Key words
Theileria annulata/Tams1/single B cell antibody分类
农业科技引用本文复制引用
张楚晗,赵帅阳,朱浩瀚,苏书晓,关贵全,罗建勋,殷宏,刘军龙..环形泰勒虫Tams1单个B细胞抗体的制备及鉴定[J].中国兽医科学,2024,54(9):1209-1215,7.基金项目
国家自然科学基金项目(32202836,31972706) （32202836,31972706）
国家重点研发计划项目(2022YFD1302102) （2022YFD1302102）
中国农业科学院科技创新工程项目(ASTIP)(CAAS-ASTIP-2021-LVRI) （ASTIP）
国家肉牛牦牛产业技术体系项目(CARS-37) （CARS-37）
甘肃省基础研究创新群体项目(22JR5RA024) （22JR5RA024）
特派团项目(22CX8NA011) （22CX8NA011）
国家寄生虫资源库项目(NPRC-2019-194-30) （NPRC-2019-194-30）
