摘要
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.
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 uhrasensitive agent for the detection of certain proteins.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2008年第5期481-487,共7页
Chinese Journal of Biochemistry and Molecular Biology
基金
深圳市科技计划项目(No.2004A110)
国家十五攻关项目(No.2003BA310A23)~~
关键词
纳米金粒子
探针
寡核苷酸
抗体
gold nanoparticle
probe
oligonucleotide
antibody
