Characterization of a polymeric adsorbed coating for DNA microarray glass slides
Characterization of a polymeric adsorbed coating for DNA microarray glass slides
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DOI：
10.1021/ac0352629
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发表时间：
2004-03-01
影响因子：
7.4
通讯作者：
Depero, LE
中科院分区：
文献类型：
--
作者：
Pirri, G;Damin, F;Depero, LE
开发了一种新的方法，将靶分子共价连接到玻璃基底（如微孔板、珠、管和显微镜载玻片）的表面上，用于与荧光靶的杂交测定。这项工作引入的创新概念是将功能性共聚物物理吸附到未衍生化的玻璃表面上，能够接枝氨基修饰的DNA分子。通过自由基共聚N，N-二甲基丙烯酰胺、N-丙烯酰氧基琥珀酰亚胺和甲基丙烯酸3-（三甲氧基甲硅烷基）丙酯共聚（DMA-NAS-MAPS）获得的聚合物非常快速地（通常在5-30分钟内）自吸附到玻璃表面上。表面层在含有各种添加剂（SDS、尿素、盐）的水性缓冲液中稳定，即使在沸腾温度下也是如此。应该强调的是，涂层是通过将载玻片浸入聚合物的稀释水溶液中形成的。因此，该过程快速、廉价、稳健且可靠，并且不需要耗时的玻璃预处理。用共聚物（DMA-NAS-MAPS）包被的载玻片可有利地用作制备低密度DNA微阵列的基底。通过X射线反射率测量来评价膜的密度和厚度，而通过功能测试来确定官能团与DNA分子的反应程度。实验表明，存在于表面上的活性基团的一半与寡核苷酸探针反应。
A new method was developed to covalently attach target molecules onto the surface of glass substrates such as microwell plates, beads, tubes, and microscope slides, for hybridization assays with fluorescent targets. The innovative concept introduced by this work is to physically adsorb onto underivatized glass surfaces a functional copolymer, able to graft amino-modified DNA molecules. The polymer, obtained by radical copolymerization of N,N-dimethylacrylamide, N-acryloyloxysuccinimide, and 3-(trimethoxysilyl)propyl methacrylate, copoly(DMA-NAS-MAPS), self-adsorbs onto the glass surface very quickly, typically in 5-30 min. The film, formed on the surface, bears active esters, which react with amino-modified DNA targets. The surface layer is stable in an aqueous buffer containing various additives (SDS, urea, salt), even at boiling temperature. It should be emphasized that the coating is formed by the immersion of glass slides in a diluted aqueous solution of the polymer. Therefore, the procedure is fast, inexpensive, robust, and reliable, and it does not require time-consuming glass pretreatments. Slides, coated with copoly(DMA-NAS-MAPS), were profitably used as substrates for the preparation of low-density DNA microarrays. The density and the thickness of the films were evaluated by X-ray reflectivity measurements whereas the extent of reaction of functional groups with DNA molecules was determined by a functional test. The experiments indicate that half of the active groups present on the surface reacts with oligonucleotide probes.