五氯酚人工抗原的合成与多克隆抗体的制备  

Synthesis of Artificial Antigen and Production of Polyclonal Antibody for Pentachlorophenol

作　　者：余宇燕[1] 唐舒雅[1] 庄惠生[1] 

机构地区：[1]东华大学环境科学与工程学院,上海200051

出　　处：《农业环境科学学报》2007年第1期314-317,共4页Journal of Agro-Environment Science

基　　金：国家自然科学基金资助项目(20277005);福建省科技厅省资助省属高校项目(2006F5054)

摘　　要：采用氯乙酸法合成五氯酚(PCP)的衍生物五氯苯氧乙酸半抗原,在此基础上,采用活化酯法合成五氯酚的人工抗原,以合成的人工抗原免疫兔子,制备了高效价的抗五氯酚多克隆抗体,抗体经纯化后,建立间接竞争荧光免疫分析新方法。分析结果表明,在优化的条件下,五氯酚检测的线性浓度范围为0.5～50!g·L-1,检测限为0.29!g·L-1,其他结构类似的酚化合物不干扰五氯酚的测定。将此法初步应用于土壤中五氯酚的快速检测,回收率在89.8%～106.5%之间,变异系数在2.85%～7.36%之间,符合农药残留分析的要求。A hapten （pentachlorophenoxiacetic acid） was synthesized by reaction of the sodium chloroacetate with sodium salts ofpentachlorophenol （PCP）. The structure of the hapten was confirmed by IR and ^1HNMR. Then artificial antigens of PCP with two kinds of proteins （BSA and OVA） were prepared by the modified active ester method. The conjugation formation was confirmed a success by UV spectrum scanning. The ratios of two kinds of conjugates （PCP-BSA and PCP-OVA） were 14：1 and 10：1, respectively. The anti-PCP polyclonal antibodies were obtained by using the artificial immune antigen to immune the New Zealand white rabbits. The antibody with high titer from the rabbit was sepa- rated from anti-serum by octanoic acid ammonium sulfate two-step precipitation method and purified by adsorption of DEAE-cellulose method. Based on these, the method of the indirect competitive fluoroimmunoassay for PCP was established successfully. The regression equation was F=24.15logC＋33.03 （r=-0.992 6） with the detection limit about 0.29 μg·L^-1. The linear detection ranged from 0.5 μg·L^-1 to 50μg·L^-1. The cross-reaction tests indicated that most structurally related phenolic compounds did not interfere with the analysis of PCP, suggesting the anti-PCP antibody had excellent recognition specificity to PCP. The method was primarily applied to detect PCP in soil samples. The recoveries ranged from 89.8% to 106.5% and coefficients of variability were from 2.85% to 7.36%. The FIA method provided a rapid, sensitive and accurate procedure for the determination of PCP in soil samples.

关 键 词：五氯酚 人工抗原 抗体制备 荧光免疫 

分 类 号：X830.2[环境科学与工程—环境工程]