磺胺二甲嘧啶单克隆抗体的制备及其鉴定  

Preparation and Identification of Monoclonal Antibody against Sulfadimidine

作　　者：王颖[1,2] 安宇[3] 李研东[2] 沈庆丰[2] 杜承[2] 柳增善[2] 罗云波[4] 

机构地区：[1]黑龙江八一农垦大学食品学院食品科学教研室,黑龙江大庆163319 [2]吉林大学教育部人兽共患病重点实验室细菌室,长春130062 [3]黑龙江八一农垦大学生命科技学院,黑龙江大庆163319 [4]中国农业大学食品学院,北京100083

出　　处：《中国生物制品学杂志》2011年第7期835-838,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金资助课题(30671762;31000790)

摘　　要：目的制备磺胺二甲嘧(Sulfadimidine,SM2)啶单克隆抗体,并对其进行鉴定。方法经小鼠腹腔免疫SM2-BSA抗原,采用常规技术进行细胞融合,间接竞争ELISA法筛选阳性杂交瘤细胞株,有限稀释法进行亚克隆,硫酸铵-辛酸法纯化抗体,并鉴定其抗体效价、杂交瘤细胞染色体数量、相对分子质量、抗体亚型、亲和力和特异性。结果筛选到1株稳定分泌抗体的杂交瘤细胞株3D7,细胞培养液上清和抗体效价分别为1∶6.4×102和1∶1.28×105;纯化后3D7抗体效价>1∶128 000,杂交瘤细胞染色体平均数为50±2对,分泌的抗体属于IgG1亚类,相对分子质量为164 280,亲和常数为6.09×108 M-1,与6种磺胺类药物及青霉素、庆大霉素和泰乐霉素交叉反应IC50大于1 mg/ml,交叉反应率小于0.01%。结论已成功制备了SM2单克隆抗体,该抗体能满足建立免疫学检测方法的需要。Objective To prepare and identify the monoclonal antibody（McAb） against sulfadimidine（SM2）.Methods BALB/c mice were immunized i.p.with SM2-BSA antigen for cell fusion by routine technique.Positive hybridoma cell strains were screened by indirect competitive ELISA,subcloned by limit dilution and determined for number of chromosome.McAb was purified by ammonium sulphate-caprylic acid method and determined for titer,relative molecular mass,subtype,affinity and specificity.Results One hybridoma cell strain stably secreting McAb,named as 3D7,was screened,and the titers of cell culture supernatant and secreted McAb were 1 ∶ 6.4 × 102 and 1 ∶ 1.28 × 105 respectively.The purified McAb 3D7 reached a titer of more than 1 ∶ 128 000.The mean number of chromosomes in the hybridoma cells was（50 ± 2） pairs.The secreted McAbs were of IgG1 subtype,of which the relative molecular mass and affinity constant were 164 280 and 6.09 × 108 M-1 respectively.All the IC50 of McAbs to six kinds of sulfa drugs,penicillin,gentamicin and tylosin were more than 1 mg/ml,and the cross reaction rate was less than 0.01%.Conclusion The McAb against SM2 was successfully prepared,which met the requirements for development of immunological detection method.

关 键 词：磺胺二甲嘧啶 抗体 单克隆 

分 类 号：R392-33[医药卫生—免疫学]