高有机溶剂耐受性的苯并(a)芘单克隆抗体制备及其免疫反应特性研究  

Preparation of monoclonal antibody to benzo(a)pyrene with high organic solvent tolerance and characterization of its immune response

作　　者：孙清 黄刚锋[3] 史亚利 时国庆 蔡亚岐[1,2] SUN Qing;HUANG Gang-Feng;SHI Ya-Li;SHI Guo-Qing;CAI Ya-Qi(School of Environment,Hangzhou Institute for Advanced Study,University of Chinese Academy of Sciences,Hangzhou 310024,China;State Key Laboratory of Environmental Chemistry and Ecotoxicology,Research Center for Eco-environmental Sciences,Chinese Academy of Sciences,Beijing 100085,China;School of Chemistry and Biological Engineering,University of Science and Technology Beijing,Beijing 100083,China)

机构地区：[1]国科大杭州高等研究院环境学院,杭州310024 [2]中国科学院生态环境研究中心环境化学与生态毒理学国家重点实验室,北京100085 [3]北京科技大学化学与生物工程学院,北京100083

出　　处：《食品安全质量检测学报》2022年第18期5895-5901,共7页Journal of Food Safety and Quality

基　　金：国家自然科学基金项目(22176198);中央高校基本科研业务费项目(FRF-BR-20-03B)。

摘　　要：目的制备适应油脂样品中苯并(a)芘[benzo(a)pyrene,BaP]检测需求的抗BaP单克隆抗体。方法利用杂交瘤单克隆抗体技术制备抗BaP的抗体,在抗体筛选过程中使用高浓度的有机溶剂(50%甲醇)来选择具有较高有机溶剂耐受能力的抗体,并利用间接竞争酶联免疫吸附法考察抗体的免疫反应特性。结果筛选获得一株对有机溶剂有较高耐受能力的、稳定分泌BaP单克隆抗体的细胞株2H10,该抗体为IgG1亚型,可耐受高浓度的甲醇、丙酮、二甲亚砜(dimethyl sulfoxide,DMSO)和二甲基甲酰胺(dimethylformamide,DMF)。60%甲醇和60%DMSO作为样品稀释液时,抗体对BaP的半抑制浓度(half maximal inhibitory concentration,IC50)分别为81.4和24.5 ng/mL,以60%DMSO作为样品稀释液,可以降低BaP抗体对其他多环芳烃化合物的交叉反应率。结论本研究获得高特异性的BaP人工抗原及高有机溶剂耐受性的抗BaP单克隆抗体。Objective To prepare monoclonal antibodies against benzo(a)pyrene(BaP)to meet the needs of BaP detection in oil samples.Methods A monoclonal antibody against BaP were prepared by hybridoma technique.A high concentration of organic solvent(50%methanol)was used in the antibody screening process to select antibodies with high tolerance to organic solvents,and the immunoreactivity characteristics of the antibody were investigated by indirect competitive enzyme-linked immunosorbent assay.Results A cell line 2H10 with high tolerance to organic solvents and stable secretion of monoclonal antibody against BaP was screened.The antibody(IgG1 isotype)could tolerate high concentrations of methanol,acetone,dimethyl sulfoxide(DMSO),and dimethylformamide(DMF).The half maximal inhibitory concentrations(IC50)of the antibody against BaP were 81.4 and 24.5 ng/mL when 60%methanol and 60%DMSO were used as the sample diluents,respectively.Using 60%DMSO as the sample diluent reduced the cross-reactivities of the BaP antibody against other polycyclic aromatic hydrocarbons compounds.Conclusion In this study,an artificial antigen with high specificity of BaP and an anti-BaP monoclonal antibody with high tolerance to organic solvents have been obtained.

关 键 词：苯并(A)芘 单克隆抗体 有机溶剂耐受 酶联免疫吸附测定 

分 类 号：R392-33[医药卫生—免疫学]