Corrie Simon R, Lawrie Gwendolyn A, Trau Matt
Centre for Nanotechnology and Biomaterials, The University of Queensland, QLD 4072, Australia.
Langmuir. 2006 Mar 14;22(6):2731-7. doi: 10.1021/la052433r.
A strategy for the production and subsequent characterization of biofunctionalized silica particles is presented. The particles were engineered to produce a bifunctional material capable of both (a) the attachment of fluorescent dyes for particle encoding and (b) the sequential modification of the surface of the particles to couple oligonucleotide probes. A combination of microscopic and analytical methods is implemented to demonstrate that modification of the particles with 3-aminopropyl trimethoxysilane results in an even distribution of amine groups across the particle surface. Evidence is provided to indicate that there are negligible interactions between the bound fluorescent dyes and the attached biomolecules. A unique approach was adopted to provide direct quantification of the oligonucleotide probe loading on the particle surface through X-ray photoelectron spectroscopy, a technique which may have a major impact for current researchers and users of bead-based technologies. A simple hybridization assay showing high sequence specificity is included to demonstrate the applicability of these particles to DNA screening.
本文提出了一种制备生物功能化二氧化硅颗粒并对其进行后续表征的策略。这些颗粒经过设计，能够制备出一种双功能材料，该材料既能（a）连接用于颗粒编码的荧光染料，又能（b）对颗粒表面进行顺序修饰以偶联寡核苷酸探针。采用了显微镜和分析方法相结合的方式，以证明用3-氨丙基三甲氧基硅烷对颗粒进行修饰会使胺基在颗粒表面均匀分布。有证据表明，结合的荧光染料与附着的生物分子之间的相互作用可忽略不计。通过X射线光电子能谱采用了一种独特的方法来直接定量颗粒表面上的寡核苷酸探针负载量，该技术可能会对当前基于微珠技术的研究人员和用户产生重大影响。包含一个显示出高序列特异性的简单杂交试验，以证明这些颗粒在DNA筛选中的适用性。
