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当前位置: 首页 > 文章 > 五氯酚人工抗原的合成与多克隆抗体的制备 农业环境科学学报 2007,26 (01) 314-317
Position: Home > Articles > Synthesis of Artificial Antigen and Production of Polyclonal Antibody for Pentachlorophenol Journal of Agro-Environment Science 2007,26 (01) 314-317

五氯酚人工抗原的合成与多克隆抗体的制备

作  者：
余宇燕;唐舒雅;庄惠生
单  位：
东华大学环境科学与工程学院
关键词：
五氯酚;人工抗原;抗体制备;荧光免疫
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。
译  名：
Synthesis of Artificial Antigen and Production of Polyclonal Antibody for Pentachlorophenol
作  者：
YU Yu-yan,TANG Shu-ya,ZHUANG Hui-sheng(College of Environment Science and Engineering,Donghua University,Shanghai 200051,China)
关键词：
pentachlorophenol;synthetic antigen;polyclonal antibody;fluoroimmunoassay
摘  要：
A hapten(pentachlorophenoxiacetic acid) was synthesized by reaction of the sodium chloroacetate with sodium salts of pentachlorophenol(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 separated 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 g·L-1.The linear detection ranged from 0.5 g·L-1 to 50 g·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.

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