多环芳烃-菲单克隆抗体制备及鉴定

孙宇;李岩松;孟宪梅;张莹;孟雷;周玉

【期刊名称】《吉林农业大学学报》

【年(卷),期】2017(039)003

【摘要】In order to rapidly and accurately detect phenanthrene (Phe) in the environment,a monoclonal antibody (McAb) against Phe was prepared,and the immunological properties of the McAb were analyzed.2-phenanthrene butanoic acid was conjugated with bovine serum albumin (BSA) and ovalbumin (OVA) for immunogen (Phe-BSA) and coating antigen (Phe-OVA) by active ester method.The conjugates were confirmed by nondenaturing agarose gel electrophoresis and ultraviolet spectrophotometry,respectively.Balb/C mice were immunized by footpad immunization method.After cell fusion,hybridoma selection and cloning,McAb against Phe was prepared.The results indicated that complete antigens of Phe were synthesized successfully.1 G7 cell line that stably secreted McAb against Phe was obtained.Titer,average affinity,IC5oand isotype of the McAb were 1 ∶ 6.4 × 105,3.29×108 L/mol,17.82 ng/mL and IgG1,respectively.The results of cross-reactivity showed that five PAHs had high CR value with the McAb,demonstrating that McAb has the potential to be used for broad spectrum assay of Phe and other homologous PAHs.%为快速、准确检测环境中的菲(Phe),研究制备了抗菲单克隆抗体,并分析了抗菲单克隆抗体的免疫学特性.采用活泼酯法将