摘要
目的制备能用于Mur抗原血清学检测的抗-Mur试剂,并将其应用于献血者标本的筛查中。方法采用偶联了载体蛋白——钥孔血蓝蛋白(KLH)的Mur抗原特异性多肽免疫BALB/c小鼠,通过淋巴细胞杂交瘤技术获得分泌抗体的细胞株,用Mur抗原阳性细胞筛选阳性克隆,利用谱细胞鉴定单克隆抗体的特异性,利用制备的单克隆抗体对1062例献血者标本的Mur抗原进行检测。结果经筛选及鉴定后共获得4个能稳定分泌抗-Mur的单克隆,均为IgM型Kappa轻链,效价分别为64、128、256和256。制备的抗-Mur单克隆抗体与日本惠赠的人源单克隆抗-Mur分别检测了1062例献血者标本,其检测结果符合率为100%。结论成功建立了能稳定分泌抗-Mur单克隆抗体的杂交瘤细胞株,可用于临床上Mur抗原的血清学快速检测。
Objective To prepare anti-Mur monoclonal antibody for serological detection of Mur antigen and apply to screening blood donors.Methods BALB/c mice were immunized with Mur antigen-specific polypeptide coupled with the carrier protein keyhole limpet hemocyanin(KLH),the cell line secreting anti-Mur was prepared by lymphocyte hybridoma technique.The positive clones were screened by Mur(+)cells,and the specificity of anti-Mur monoclonal antibody was identified by the antibody identification panel cells.Finally,the prepared monoclonal anti-Mur was used to detect 1062blood donor samples.Results Through screening and identification,four hybridoma cell lines that could stably secrete anti-Mur were obtained,all of which were Ig M kappa chain with titers of 64,128,256 and 256,respectively.The prepared anti-Mur monoclonal antibody and the human monoclonal anti-Mur donated in the Japanese were used to detect 1062 blood donors respectively,and the results were consistent(100%).Conclusion Hybridoma cell lines that could stably secrete anti-Mur monoclonal antibody were successfully established,which could be used for rapid serological detection of Mur antigen in clinic.
作者
魏玲
王贞
朱思颖
骆宏
罗广平
姬艳丽
WEI Ling;WANG Zhen;ZHU Si⁃ying;LUO Hong;LUO Guang⁃ping;JI Yan⁃li(Institute of Clinical Blood Transfusion,Guangzhou Blood Center,Guangzhou,Guangdong 510095,China)
出处
《热带医学杂志》
CAS
2022年第7期929-932,937,共5页
Journal of Tropical Medicine
基金
国家自然科学基金(81901492)
广州市科技计划项目(201904010048)
广州市医学重点实验室项目
