克百威人工抗原合成及其免疫应用
Synthesis and Immune Application of Artificial Antigen of Carbofuran

研究克百威半抗原4-［〔（2，3-二氢-2，2-二甲基-7-苯并呋喃基氧）羰基〕氨基］丁酸（BFNB）的新合成方法，以避免传统三光气合成法的危害。以呋喃酚为起始物，通过酯化反应和还原反应最终合成半抗原BFNB，利用质谱和核磁进行表征，结果证明半抗原BFNB合成成功。利用活性酯法将BFNB分别与牛血清白蛋白（BSA）和卵清蛋白（OVA）偶联得到免疫原和包被原，紫外扫描计算偶联比分别为17∶1和8∶1；将免疫原免疫小鼠制备多克隆抗体，间接竞争酶免疫检测方法（icELISA）表明其半数抑制质量浓度（IC50）为0.096 μg/mL，定量检测线性（IC20~IC80）为0.023~0.403 μg/mL。该研究所制备的抗原为进一步研制抗克百威单克隆抗体奠定基础。
To explore new method for the synthesis of 4-［〔(2,3-Dihydro-2,2-dimethyl-7-benzofuranyloxy)carbonyl〕-amino］butanoic acid (BFNB), and avoided the damage of traditional carbonyl chloride synthesis technology. Hapten BFNB was synthesized by ester reaction with benzofuranol as raw material and was characterized by mass spectrometry and H NMR.The results showed that the structure of product was consistent with the target compound. Artificial antigens were prepared by active ester method, and then were identified by UV spectrophotometric and calculated the coupling ratio. According to the UV, the artificial antigen of BFNB-BSA and BFNB-OVA were synthesized successfully and the coupling ratios were 17∶1 and 8∶1, respectively. The BSA conjugates were used to immunize BALB/c mice for preparation of antibody while OVA conjugates were used as coating antigen. Indirect competitive ELISA methods were developed with IC50 value of 0.096 μg/mL and the linear range is 0.023-0.403 μg/mL. The preparation of antigen laid ranged foundation for further preparation of monoclonal antibody to carbofuran.