中国农业科技导报2014，Vol.16Issue(3)：143-149,7.DOI:10.13304/j.nykjdb.2013.548
一种新的抗多环芳烃多克隆抗体的制备
Preparation of a New Polyclonal Antibody Based on PAHs Immunogen
摘要
Abstract
In order to exploit the polycyclic aromatic hydrocarbons (PAHs) ELISA test kit for rapidly detecting PAHs in different environmental samples,the polyclonal antibody (pAb) against pyrene was prepared.The hapten (1-Pyrenebutyric acid,1-PBA) was conjugated covalently to the carrier proteins including bovine serum albumin (BSA) and ovalbumin (OVA)to prepare immunogen and coating antigen via amide bonds by the active ester method.Antiserum was raised against the conjugate PBA-BSA by stimulating immune responses in rabbits.The specific pAb against PAHs was obtained by affinity purification and developing an indirectly competitive enzyme-linked immunosorbent assay (ic-ELISA).A standard curve for pyrene was established to detect the specificity of antibody.The titer of compound antiserum was 1∶102 400.The average IC50 value was 0.06 mg/L,and the detection limit at 10％ inhibition (IC90) was 0.01 mg/L.The pAb had the highest affinity with 4 ～ 6 ring PAHs by determining crossreactivity of 16 PAHs.Further,the additive of less than 40％ methanol in the test sample has no significant effect on ELISA.The PAHs polyclonal antibody and its characterization are necessary for technical reserves in future development and improvement of the PAHs ELISA test kit.关键词
多环芳烃/芘/多克隆抗体/酶联免疫吸附分析法Key words
PAHs / pyrene/ polyclonal antibody/ ELISA分类
生物科学引用本文复制引用
许丽,赵鲁,程言君,孙恺璇,赵可卉..一种新的抗多环芳烃多克隆抗体的制备[J].中国农业科技导报,2014,16(3):143-149,7.基金项目
Special Item for Reform and Development of Environmental Protection Research Institute of Light Industry (2012A-7) supported. （2012A-7）
