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在涂有聚丙烯酸的玻璃上接枝聚乙二醇以形成抗蛋白质表面。

Grafting of poly(ethylene glycol) onto poly(acrylic acid)-coated glass for a protein-resistant surface.

作者信息

Wazawa Tetsuichi, Ishizuka-Katsura Yoshiko, Nishikawa So, Iwane Atsuko Hikikoshi, Aoyama Shigeru

机构信息

OMRON-Endowed Chair in Nano Optical Devices, and Soft Biosystem Laboratory, Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita, Osaka 565-0871 Japan.

出版信息

Anal Chem. 2006 Apr 15;78(8):2549-56. doi: 10.1021/ac052102j.

Abstract

The surface of solid glass supports for samples in optical microscopy and for biosensors needs to be protein-resistant. A coating of a poly(ethylene glycol) monomethyl ether (mPEG) on the surface of the glass is one promising method for preventing the nonspecific adsorption of proteins. In this study, we have developed a novel technique for achieving an optimal coverage of a glass surface with mPEG to prevent protein adhesion. A clean glass substrate previously treated with (3-aminopropyl)dimethylethoxysilane (APDMES) was treated sequentially with poly(acrylic acid) and subsequently a primary amine derivative of mPEG in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. The resultant glass surface was demonstrated to be highly protein-resistant, and the adsorption of bovine serum albumin decreased to only a few percentage points of that on a glass surface treated with APDMES alone. Furthermore, to extend the present method, we also prepared a glass substrate on which biotinylated poly(ethylene glycol) was cografted with mPEG, and biotinylated myosin subfragment-1 (biotin-S1) was subsequently immobilized on this substrate by biotin/avidin chemistry. Actin filaments were observed to glide on the biotin-S1-coated glass surface in the presence of ATP, and thus, the method is capable of immobilizing the protein specifically without any loss in its biological function.

摘要

用于光学显微镜样品和生物传感器的固体玻璃载体表面需要具备抗蛋白性。在玻璃表面涂覆聚乙二醇单甲醚(mPEG)是防止蛋白质非特异性吸附的一种很有前景的方法。在本研究中,我们开发了一种新技术,可实现玻璃表面mPEG的最佳覆盖,以防止蛋白质粘附。先用(3-氨丙基)二甲基乙氧基硅烷(APDMES)处理过的干净玻璃基板,依次用聚丙烯酸处理,随后在1-乙基-3-(3-二甲基氨基丙基)碳二亚胺存在的情况下用mPEG的伯胺衍生物处理。结果表明,所得玻璃表面具有高度抗蛋白性,牛血清白蛋白的吸附量仅降至单独用APDMES处理的玻璃表面吸附量的几个百分点。此外,为扩展本方法,我们还制备了一种玻璃基板,在其上将生物素化聚乙二醇与mPEG共接枝,随后通过生物素/抗生物素蛋白化学将生物素化肌球蛋白亚片段-1(生物素-S1)固定在该基板上。在ATP存在的情况下,观察到肌动蛋白丝在生物素-S1包被的玻璃表面上滑动,因此,该方法能够特异性地固定蛋白质,而不会使其生物学功能有任何损失。

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