小分子阿特拉津和罂粟碱检测的免疫芯片技术研究

Research on Protein Chip Technologies for the Detection ofAtrazine and Papaverine

采用蛋白芯片竞争法对小分子半抗原的污染物进行检测。在获得特异性抗体的前提下,首先将阿特拉津半抗原进行了衍生化,然后将该衍生物和氨基罂粟碱分别与载体蛋白质卵清蛋白(OVA)进行偶联。实验证明新合成的完全抗原能够与其相应抗体发生特异性的结合。实验还对蛋白芯片检测阿特拉津进行了条件优化,其抗体固定化时间为2h,用卵清蛋白为封闭液的封闭时间为1h,样品稀释液pH值为8. 0。并对阿特拉津及罂粟碱进行了定性、定量实验,结果表明:荧光信号强度随待测物浓度的降低而增强,有一定的线性趋势,阿特拉津检出限为0. 001mg/L,罂粟碱检出限为0. 01mg/L。

Protein chip is a rapidly expanding technology for large-scale, high-throughput protein assay after the. genechip. A competitive inhibition method for screening of poisonous chemical molecule haptens was adopted due to the aiming contaminant characteristic of small molecule. Having obtained antibodies, the derivatives of pesticide atrazine and parathion were successfully synthesized, then the derivatives and amino-papaverine were conjugated to protein ovalbumin(OVA), used as antigens which proved subsequently high specificity against its corresponding antibody. The detection of atrazine and papaverine was performed qualitatively and quantitatively under the optimized conditions which includes antibody immobilized time (2h), blocking buffer containing OVA and blocking time (1 h), sample buffer pH value ( pH 8. 0). The results demonstrate that fluorescence intensity increased along with the decreasing concentration of analytes. And then, linear trend obtained. The limit of detection of atrazine and papaverine is 0.001 and. 0. 01 mg/L respectively.