重庆理工大学学报2024，Vol.38Issue(15)：291-297,7.DOI:10.3969/j.issn.1674-8425(z).2024.08.034
布鲁氏菌LPS单克隆抗体的制备与鉴定
Preparation and identification of monoclonal antibodies against Brucella abortus LPS
摘要
Abstract
A monoclonal antibody specific to Brucella LPS is prepared to provide effective reagents for the establishment of brucellosis serology and novel immunosensor assays.The naturally extracted Brucella LPS antigen is injected intraperitoneally into BALB/c mice using the long-range immunization method.After five times of routine immunizations and one time of enhanced immunization,cell fusion of mouse splenocytes and myeloma cells (SP2/0) is performed using PEG1500 as a pro-fusion agent;hybridoma cells are screened by indirect enzyme-linked immunosorbent assay (iELISA).Our results show three positive cell lines with stable monoclonal antibody secretion are named 2-4A,3-4E and 5-10B,and after the identification of subtypes,3-8E and 2-4A are IgG1 subtypes and 5-10B is IgG2b subtype.The 5-10B cell line is employed to prepare ascites,which is purified by Protein A column.After purification,the monoclonal antibody reaches electrophoretic purity,and its concentration reaches up to 1.4318 mg·mL-1 as determined by BCA method,and the monoclonal antibody is purified to a titer of 1∶512000.The indirect ELISA results show the prepared antibody has good specificity.The highly specific and active monoclonal antibody prepared in this study is expected to lay the foundation for the subsequent diagnosis of brucellosis.关键词
布鲁氏菌/LPS/细胞融合/单克隆抗体/生物特性鉴定Key words
Brucella abortus/LPS/cell fusion/monoclonal antibody/biological characterization分类
农业科技引用本文复制引用
廖倩,向梦玲,凌洪权,董春霞,骆璐,吴胜昔,曾政..布鲁氏菌LPS单克隆抗体的制备与鉴定[J].重庆理工大学学报,2024,38(15):291-297,7.基金项目
重庆理工大学研究生创新基金项目(gzlcx20233385) （gzlcx20233385）
丰都肉牛疾病诊断技术开发项目"肉牛三种重大疫病快速诊断试剂的研发"(2023Q23) （2023Q23）
重庆市现代农业产业技术体系创新团队项目(CQMAITS202314) （CQMAITS202314）
