Dendane Nabil, Hoang Antoine, Guillard Ludovic, Defrancq Eric, Vinet Françoise, Dumy Pascal
Département de Chimie Moléculaire - UMR CNRS 5250, ICMG FR2607, Université Joseph Fourier, BP 53, 38041 Grenoble Cedex 9, France.
Bioconjug Chem. 2007 May-Jun;18(3):671-6. doi: 10.1021/bc060254v. Epub 2007 Mar 10.
The efficient surface patterning of oligonucleotides was accomplished onto the inner wall of fused-silica capillary tubes as well as on the surface of glass slides through oxime bond formation. The robustness of the method was demonstrated by achieving the surface immobilization of up to three different oligonucleotide sequences inside the same capillary tube. The method involves the preparation of surfaces grafted with reactive aminooxy functionalities masked with the photocleavable protecting group, 2-(2-nitrophenyl) propyloxycarbonyl group (NPPOC). Briefly, NPPOC-aminooxy silane 1 was prepared and used to silanize the glass surfaces. The NPPOC group was cleaved under brief irradiation to unmask the reactive aminooxy group on surfaces. These reactive aminooxy groups were allowed to react with aldehyde-containing oligonucleotides to achieve an efficient surface immobilization. The advantage associated with the present approach is that it combines the high-coupling efficiency of oxime bond formation with the convenience associated with the use of photolabile groups. The present strategy thus offers an alternative approach for the immobilization of biomolecules in the microchannels of "labs on a chip" devices.
通过肟键形成,可将寡核苷酸高效地图案化到熔融石英毛细管内壁以及载玻片表面。通过在同一根毛细管内实现多达三种不同寡核苷酸序列的表面固定,证明了该方法的稳健性。该方法包括制备接枝有被光可裂解保护基团2-(2-硝基苯基)丙氧基羰基(NPPOC)掩盖的反应性氨氧基官能团的表面。简而言之,制备了NPPOC-氨氧基硅烷1并用于硅烷化玻璃表面。在短暂照射下NPPOC基团被裂解,以暴露表面上的反应性氨氧基基团。使这些反应性氨氧基基团与含醛寡核苷酸反应,以实现高效的表面固定。本方法的优点在于它将肟键形成的高偶联效率与使用光不稳定基团的便利性相结合。因此,本策略为在“芯片实验室”装置的微通道中固定生物分子提供了一种替代方法。