呋喃丹多克隆抗体的制备与初步应用

以呋喃丹为原料，在强碱性(pH>12)下水解生成呋喃酚，与对硝基苯氯甲酸酯和6-氨基己酸反应，合成呋喃丹半抗原，即6-［［(2,3-二氢-2,2-二甲基-7-苯并呋喃基氧)羰基］氨基］己酸(BFNH). 通过活性酯法将半抗原与载体BSA偶联制备的呋喃丹人工免疫抗原免疫家兔获得了抗呋喃丹多克隆抗体，免疫抗原BFNH-BSA和包被抗原BFNH-OVA的结合比分别为18∶1和5∶1.采用辛酸-硫酸铵二步沉淀法纯化兔Ⅱ抗体，抗体最高效价为5.12×106，重、轻链分子量分别约为55和22 ku.通过间接竞争ELISA鉴定抗体质量，测得在10-7～0.01 mg/mL范围内，抑制率与标准农药浓度的线性关系显著，呋喃丹残留最低检测限为10-7 mg/mL，与5种结构相似的氨基甲酸酯类的交叉反应率低于10%. 并初步应用于果蔬样品呋喃丹残留的快速检测. 

Production and Initial Application of Carbfuran Polyclonal Antibody

A hapten 6-［［(2, 3-dihydro-2, 2-dimetyl-7-benzofuranyloxy) carbonyl］ amino］ hexanoic acid (BFNH) completely preserving the carbamate group of carbofuran was synthesized with 2, 3-dihydro-2, 2-dimrthyl-7-benzofuranol and aminocaproic acid, which was activated with 4-nitrophenyl chloroform-mate. The BFNH hapten was attached to bovine serum album (BSA) by utilizing the modified active ester method to synthesize antigen BFNH-BSA. The BFNH-BSA was used as immunogen, with which an anti-carbofuran polyclonal antibody (pAb) was successfully obtained by immunization of rabbits. The coupling ratios of BFNH-BSA and BFNH-OVA were 18∶1 and 5∶1, respectively. The highest titer of the pAb was 5.12×106 by purifying rabbit II with the caprylid acid method. The linear concentrations ranged from 10-7 mg/mL to 0.01 mg/mL and the detection limit was 10-7 mg/mL for the determination of carbofuran by a competitive indirect ELISA procedure. The cross-reactivities of five oxaminic acidesters were lower than 10%. And the pAb was applied initially to detect carbofuran reside in fruit and vegetables.