摘 要:本研究采用抗原决定簇法,在硅烷化的玻片表面合成制备出对缩胆囊素神经肽具有特异性识别能力的分子印迹膜。优化了印迹膜合成制备条件,并用扫描电镜和红外光谱对其进行表征,通过吸附平衡实验评价印迹膜的吸附容量及选择性。基于此分子印迹膜,研究建立了固相竞争-荧光免疫分析方法,利用荧光倒置显微镜-CCD图像分析系统对人脑脊液中缩胆囊素神经肽进行定量分析。实验结果表明所制备的分子印迹膜具有特异性强和可重复利用的优点,在生物体液中缩胆囊素神经肽分析测定上具有良好的应用价值。
关键词:缩胆囊素;神经肽;分子印迹聚合物;竞争免疫
Preparation of Cholecystokinin Neuropeptides Molecularly Imprinted Membrane and Its Application in Fluorescence Immunoassay
Abstract:A molecularly imprinted polymer(MIP) membrane for highly selective recognition of cholecfstokinin(CCK) neuropeptides was synthesized on glass slide by epitope imprinting technique.The polymerization conditions were investigated and optimized.The synthesized MIP membrane was characterized by scanning electron microscopy and infrared spectroscopy,and its adsorption capacity and selectivity were evaluated using equilibrium adsorption tests.An imaging assay,which is analogous to solid-phase competitive immunoassay,was developed using this MIP membrane as the support for fluorescence reaction and used for quantification of CCK neuropeptides in human cerebrospinal fluids by inverted fluorescence microscope-CCD imaging system.The limit of detection for CCK neuropeptides was 8.91 pmol/L.The economical and reusable properties of this MIP membrane suggest its potential for the assay of CCK neuropeptides in biological fluids.
Keywords:Cholecystokinin;Neuropeptides;Molecularly imprinted polymer;Competitive immunoassay
分析科学