1-芘丁酸单克隆抗体的制备及免疫学性能鉴定  

Preparation and immunological characteristics identification of monoclonal antibody to 1-Pyrenebutyric acid

作　　者：胡骁飞[1] 孔蒙蒙 邢广旭[1] 邢云瑞 孙亚宁[1] 张改平[2] HU Xiao-Fei;KONG Meng-Meng;XING Guang-Xu;XING Yun-Rui;SUN Ya-Ning;ZHANG Gai-Ping(Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China)

机构地区：[1]河南省农业科学院动物免疫学重点实验室,郑州450002 [2]河南农业大学,郑州450002

出　　处：《中国免疫学杂志》2020年第9期1114-1118,共5页Chinese Journal of Immunology

基　　金：国家“十二五”科技支撑计划课题(2014BAD13B05);中国博士后基金项目(2018M632778)项目资助。

摘　　要：目的:拟制备灵敏、特异的1-芘丁酸(PBA)单克隆抗体并对其免疫学性能进行鉴定。方法:碳二亚胺法(EDC)制备人工抗原PBA-BSA和PBA-OVA。PBA-BSA免疫BALB/c小鼠,选择血清效价高、灵敏度良好的小鼠进行细胞融合并筛选出分泌抗PBA单克隆抗体的阳性杂交瘤细胞株,体内诱生腹水法批量制备抗体并对其免疫学性能进行鉴定。结果:筛选出能稳定分泌PBA单克隆抗体的阳性杂交瘤细胞株10C2A6,单克隆抗体效价为1∶2.56×10^5,IC50为0.86 ng/ml,亚型为IgG2b型,亲和常数为1.96×10^9 L/mol,与芘、1-芘甲醛和1-芘甲醇的交叉反应率分别为1.74%、1.42%和0.63%,与其他物质交叉反应率均小于0.05%。结论:本实验成功制备出了灵敏、特异的PBA单克隆抗体,并为其免疫学检测方法的建立奠定了基础。Objective:To prepare monoclonal antibodies against 1-Pyrenebutyric acid(PBA) with good sensitivity,high specificity and to indentify the immunological characteristics.Methods:Artificial antigens PBA-BSA and PBA-OVA were prepared by carbodiimide method.The BALB/c mice were immunized with PBA-BSA,mice with highest serum titer and good sensitivity was selected for cell fusion.Hybridoma cell lines secreting anti-PBA monoclonal antibodies were screened.PBA monoclonal antibodies were obtained by inducing ascites in vivo,and then the immunological characteristics of which was identified.Results:A hybridoma cell line that secreting monoclonal antibodies against PBA was screened,which named as 10 C2 A6.The titer of the monoclonal antibody(mAb) was 1∶2.56×10^5,the IC50 was 0.86 ng/ml,the subtype was IgG2 b,the affinity constant(Ka) was 1.96×10^9 L/mol,the cross reaction rates with pyrene,1-pyrenecarboxaldehyde and 1-pyrenemethanol were 1.74%,1.42%,0.63%,respectively,and the cross-reaction rate with other substances was less than 0.05%.Conclusion:In this experiment,PBA-mAb with high sensitivity,specificity and affinity was prepared,which provided technical support for the immunological rapid detection technology of PBA.

关 键 词：1-芘丁酸 人工抗原 细胞融合 杂交瘤细胞 单克隆抗体 

分 类 号：R155[医药卫生—营养与食品卫生学] R392[医药卫生—公共卫生与预防医学]