Sitsanidis Efstratios D, Schirmer Johanna, Lampinen Aku, Mentel Kamila K, Hiltunen Vesa-Matti, Ruokolainen Visa, Johansson Andreas, Myllyperkiö Pasi, Nissinen Maija, Pettersson Mika
Department of Chemistry, Nanoscience Center, University of Jyväskylä P. O. Box 35, FI-40014 JYU Finland
Department of Physics, Nanoscience Center, University of Jyväskylä P. O. Box 35, FI-40014 JYU Finland.
Nanoscale Adv. 2021 Feb 22;3(7):2065-2074. doi: 10.1039/d0na01028f. eCollection 2021 Apr 6.
An approach for controlled protein immobilization on laser-induced two-photon (2P) oxidation patterned graphene oxide (GO) surfaces is described. Selected proteins, horseradish peroxidase (HRP) and biotinylated bovine serum albumin (b-BSA) were successfully immobilized on oxidized graphene surfaces, non-covalent interactions, by immersion of graphene-coated microchips in the protein solution. The effects of laser pulse energy, irradiation time, protein concentration and duration of incubation on the topography of immobilized proteins and consequent defects upon the lattice of graphene were systemically studied by atomic force microscopy (AFM) and Raman spectroscopy. AFM and fluorescence microscopy confirmed the selective aggregation of protein molecules towards the irradiated areas. In addition, the attachment of b-BSA was detected by a reaction with fluorescently labelled avidin-fluorescein isothiocyanate (Av-FITC). In contrast to chemically oxidized graphene, laser-induced oxidation introduces the capability for localization on oxidized areas and tunability of the levels of oxidation, resulting in controlled guidance of proteins by light over graphene surfaces and progressing towards graphene microchips suitable for biomedical applications.
本文描述了一种在激光诱导双光子（2P）氧化图案化氧化石墨烯（GO）表面进行可控蛋白质固定的方法。通过将涂覆有石墨烯的微芯片浸入蛋白质溶液中，利用非共价相互作用，成功地将选定的蛋白质——辣根过氧化物酶（HRP）和生物素化牛血清白蛋白（b-BSA）固定在氧化石墨烯表面。通过原子力显微镜（AFM）和拉曼光谱系统地研究了激光脉冲能量、照射时间、蛋白质浓度和孵育时间对固定蛋白质形貌以及石墨烯晶格上随之产生的缺陷的影响。AFM和荧光显微镜证实了蛋白质分子向照射区域的选择性聚集。此外，通过与荧光标记的抗生物素蛋白-异硫氰酸荧光素（Av-FITC）反应检测到了b-BSA的附着。与化学氧化石墨烯不同，激光诱导氧化引入了在氧化区域定位的能力以及氧化水平的可调性，从而实现了光对石墨烯表面蛋白质的可控引导，并朝着适用于生物医学应用的石墨烯微芯片发展。
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