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首页> 外文会议>Conference on Biomedical Applications of Micro-and Nanoengineering, Dec 16-18, 2002, Melbourne, Australia >SURFACE CHARACTERIZATION OF OLIGONUCLEOTIDES IMMOBILIZED ON POLYMER SURFACES
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SURFACE CHARACTERIZATION OF OLIGONUCLEOTIDES IMMOBILIZED ON POLYMER SURFACES

机译：固定在聚合物表面上的寡核苷酸的表面表征

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摘要

The immobilization and hybridization of amino-terminated oligonucleotide strands to cyclo-olefin-copolymer (COC) and polycarbonate (PC) surfaces have been investigated for potential application in micro-PCR devices. The oligonucleotides were covalently bound to the plasma-treated COC and PC surfaces via an N-hydroxy-sulfosuccinimide (NHSS) intermediate. Analysis by AFM showed that the oligonucleotides were present on the surfaces as lumps, and that the size, both vertically and laterally, of these lumps on the COC surface was larger compared to the PC surface. The immobilization efficiency of the former was also higher (15.8 x 10~(12) molecules / cm~2) compared to the latter (3.3 x 10~(12) molecules / cm~2). The higher efficiency of the COC surface is attributed to the more effective NHSS-functionalization and its higher surface roughness. Subsequent hybridization doubled the height of the lumps, while the lateral dimensions remained essentially unchanged. This is explained in terms of organization of the long probe strands used on the surface as flexible, coil-like polymer chains, which allow the complementary oligonucleotides to bind and increase the height of the lumps. The AFM frictional images showed that the hybridization had the effect of reversing hydrophilicity of the oligonucleotide lumps from being more hydrophilic to more hydrophobic, consistent with the hydrophilic bases of the probe strands being shielded as a result of hybridization.
机译：已经研究了氨基末端寡核苷酸链与环烯烃共聚物（COC）和聚碳酸酯（PC）表面的固定化和杂交在微PCR装置中的潜在应用。寡核苷酸通过N-羟基磺基琥珀酰亚胺（NHSS）中间体与经等离子体处理的COC和PC表面共价结合。通过AFM分析显示寡核苷酸以块状存在于表面上，并且与PC表面相比，COC表面上这些块状在垂直和横向上的大小都更大。前者的固定效率（15.8 x 10〜（12）分子/ cm〜2）也比后者（3.3 x 10〜（12）分子/ cm〜2）高。 COC表面的效率更高归因于更有效的NHSS-官能化及其更高的表面粗糙度。随后的杂交使团块的高度加倍，而横向尺寸基本上保持不变。这是根据在表面上使用的长探针链作为柔性的，盘绕状的聚合物链的组织来解释的，该链允许互补的寡核苷酸结合并增加团块的高度。 AFM摩擦图像显示，杂交具有使寡核苷酸团块的亲水性从更亲水变为更疏水的作用，这与杂交所屏蔽的探针链的亲水碱基一致。

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