| 孙 清,黄刚锋,史亚利,时国庆,蔡亚岐.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[J].食品安全质量检测学报,2022,13(18):5895-5901 |
| 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 |
| 投稿时间：2022-06-30 修订日期：2022-09-03 |
| DOI： |
| 中文关键词: 苯并(a)芘 单克隆抗体 有机溶剂耐受 酶联免疫吸附测定 |
| 英文关键词:benzo(a)pyrene monoclonal antibody organic solvent tolerance enzyme-linked immunosorbent assay |
| 基金项目: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) |
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| Author | Institution |
| SUN Qing | School of Environment, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences；State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-environmental Sciences, Chinese Academy of Sciences；School of Chemistry and Biological Engineering, University of Science and Technology Beijing |
| HUANG Gang-Feng | School of Chemistry and Biological Engineering, University of Science and Technology Beijing |
| SHI Ya-Li | School of Environment, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences；State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-environmental Sciences, Chinese Academy of Sciences |
| SHI Guo-Qing | School of Chemistry and Biological Engineering, University of Science and Technology Beijing |
| CAI Ya-Qi | School of Environment, Hangzhou Institute for Advanced Study, University of Chinese Academy of Sciences；State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-environmental Sciences, Chinese Academy of Sciences |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| 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. |
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