抗体和寡核苷酸双标记纳米金生物探针的制备及性能分析

基于纳米金粒子与抗体静电吸附作用,与硫醇修饰的寡核苷酸共价结合,建立一种新的双标记纳米金生物探针的制备方法.通过透射电镜（TEM）、紫外光谱、斑点免疫金渗滤法、免疫金银染色光镜观察法、荧光标记法等检测探针表征,及表面抗体活性情况和寡核苷酸的覆盖率，同时采用变性聚丙烯酰胺凝胶电泳（PAGE）检测寡核苷酸的存在.结果表明，纳米金粒子同时连接抗体和寡核苷酸后生物性能良好，且每个纳米金粒子（10±3）nm表面可覆盖寡核苷酸(92±20)条，双标记纳米金生物探针的制备具有简捷、稳定的特点.可作为一种新型探针应用于超微量蛋白质检测.

Preparation and Characterization of Dual-labeled Gold Nanoparticle Probes withOligonucleotides and Antibodies

We experimented the preparation of the novel gold nanoparticle probes that labeled with both antibodies and thiol-terminated oligonucleotides The morphology and spectroscic properties of these probes were evaluated by transmission electron microscopy (TEM) and UV-vis spectra analyses. The degrees of antibody coating and the coverage of the oligonucleotides were analyzed by dot-immunogold filtration assay, immunogold-silver staining, fluorospectrophotometry. The oligonucleotide conjugation to the surface of （10±3）nm sized gold nanoparticles was determine to be (92±20) oligonucleotides per nanoparticle by PCR-PAGE techniques. With the high stability and ease of preparation, our dual-labeled gold nanoparticle probes could be attractive to researchers as a versatile and ultrasensitive agent for the detection of certain proteins.