摘要
为了建立有效地克隆分泌抗铜抗体杂交瘤细胞的方法,试验用不同浓度的甲基纤维素半固体培养基法培养杂交瘤细胞。结果表明:培养杂交瘤细胞的甲基纤维素最佳浓度为1.6%;获得多株阳性细胞,其中阳性细胞株3B1-C5的染色体数在100个以上,腹水抗体效价达到1∶102 400,其分泌的抗体属IgM亚类,轻链为Kappa型;经间接ELISA方法检测,连续培养单细胞系93 d,3B1-C5细胞株能较稳定分泌抗体。说明含1.6%甲基纤维素的半固体培养法适合克隆分泌抗铜抗体的杂交瘤细胞。
Toestablish the efficient cloning method of hybridoma cells that secreted antibodies against copper,the most appropriate concentration of methylcellulose in semi-solid selection medium was 1.6% for culturing the hybridoma cells that secreted antibodies against copper confirmed by the comparison testwith different methylcellulose concentration.Several positive hybridoma cell lines were obtainedwith semi-solid medium containing 1.6% methylcellulose.One of positive hybridoma cell line was named 3B1-C5.The chromosome number of 3B1-C5 were above 100.The ascites fluid was produced in Balb/c mice by intraperitoneal injection of cells of 1×106 3B1-C5.The titers of ascites fluid was up to 1∶102 400.The subtype of secreted antibody from 3B1-C5 was IgM,Kappa type.The result of indirect competitive ELISA suggested that the antibody secretion of 3B1-C5 was stable after continuous culturing above 3 months.The results of experiment indicated that the method which clone cells in semi-solid medium containing 1.6% methylcellulose was prefered to clonethe hybridoma cells secreting antibodies against copper.This cloning method is a potential method due to simple,with low cost higherpositive ratio of selectedclones,and easy-performed in the monoclonal antibody selection.
出处
《黑龙江畜牧兽医》
CAS
北大核心
2011年第12期13-16,共4页
Heilongjiang Animal Science And veterinary Medicine
基金
"863"国家高技术研究发展计划项目(2007AA10Z439)
国家自然科学基金项目(30671205)
北京市科委课题项目(Z09090501040901)
北京市农林科学院科技创新能力建设专项(KJCX201101010)
关键词
铜
杂交瘤细胞
甲基纤维素
克隆
抗体
鉴定
copper
hybridoma cells
methylcellulose
cloning
antibody
identification
