[关键词]
[摘要]
为建立一种快速检测双酚A(BPA)的方法,本论文建立以胶体金为基础的免疫层析法(GICA)。首先通过BPA人工抗原免疫Balb/c小鼠,然后利用正常的杂交瘤技术制备BPA的单克隆抗体,所制备的抗体经过辛酸-硫酸铵(CA-AS)沉淀法纯化后并鉴定为IgG2b亚型。胶体金是由柠檬酸三钠还原氯金酸而制得,通过紫外扫描光谱和透射电镜检测,结果表明胶体金在530 nm有单一吸收峰、粒径约为30 nm。用其标记BPA单克隆抗体,制备金标抗体。本研究所制备的胶体金免疫层析试纸条的检测限为100 ng/mL,检测时间为10 min,交叉反应表明,试纸条和BPA的结构类似物双酚酸、任基酚和辛基酚无交叉反应,有良好的再现性。该方法能够方便、快捷、有效的检测环境中的BPA,免疫层析试纸的研制为BPA进行现场快速的检测奠定了基础。
[Key word]
[Abstract]
To establish a rapid method for detection of Bisphenol A (BPA), colloidal gold-based immune chromatographic assay (GICA) was used. After using artificial antigen of BPA to immune Balb/C mice, monoclonal antibodies against BPA were prepared by normal hybridoma technology. The monoclonal antibody was purified by precipitation of Caprylic Acid-Ammonium Sulfate (CA-AS) and it was identified as IgG2b subtypes. Colloidal gold was prepared through the reduction of gold chloride by sodium citrate acid, UV-Vis spectrum and transmission electron microscope (TEM) were performed, the results indicated that the colloidal gold had a single 530 nm absorption peak and its particle size was about 30 nm. The colloidal gold-antibody was produced by monoclonal antibody of BPA labeled with colloidal gold. In this study, the GICA sensitivity was about 100 ng/mL and the detection time was about 10 min. The cross-reactions showed that there was no cross-reaction between test strip and BPA analogues, but had good reproducibility. This method was convenient, rapid and effective for detecting BPA, and the development of GICA was applicable for the rapid determination of BPA in the field.
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[基金项目]
国家自然科学基金资助项目(41071176;41271340)