Ameringer Thomas, Hinz Michael, Mourran Claudia, Seliger Hartmut, Groll Juergen, Moeller Martin
Deutsches Wollforschungsinstitut an der RWTH Aachen e.V., Pauwelsstrasse 8, 52074 Aachen, Germany.
Biomacromolecules. 2005 Jul-Aug;6(4):1819-23. doi: 10.1021/bm0500695.
In this study, star PEG coatings on glass substrates have been used as support material for oligonucleotide microarrays. These coatings are prepared from solutions of six armed star shaped prepolymers that carry reactive isocyanate endgroups. As described earlier, such films prevent the adsorption of proteins and the adhesion of cells but can easily be functionalized for specific biological recognition. Here we used the high functionality of these coatings for the covalent immobilization of amino terminated 20mer oligonucleotides, both by microcontact printing and spotting techniques. The permanent immobilization of fluorescently labeled DNA as well as hybridization of 20mer oligonucleotides have been monitored by fluorescence microscopy. The hybridization efficiency as determined by fluorescence intensity varied from 30% to 80% depending on the way of layer preparation. The direct spotting without additional activation and blocking steps of the surface demonstrates the potential of star PEG coatings as ultrathin surface modification for microarrays.
在本研究中,玻璃基板上的星形聚乙二醇(PEG)涂层已被用作寡核苷酸微阵列的支撑材料。这些涂层由带有反应性异氰酸酯端基的六臂星形预聚物溶液制备而成。如前所述,此类薄膜可防止蛋白质吸附和细胞黏附,但可轻松进行功能化处理以实现特定的生物识别。在此,我们通过微接触印刷和点样技术利用这些涂层的高功能性实现氨基末端20聚体寡核苷酸的共价固定。通过荧光显微镜监测了荧光标记DNA的永久固定以及20聚体寡核苷酸的杂交情况。根据荧光强度确定的杂交效率在30%至80%之间变化,具体取决于层制备方式。在不进行表面额外活化和封闭步骤的情况下直接点样,证明了星形PEG涂层作为微阵列超薄表面修饰的潜力。