Analysis Center and Department of Chemistry, Tsinghua University, Beijing 100084, PR China.
Biosens Bioelectron. 2011 Jul 15;26(11):4497-502. doi: 10.1016/j.bios.2011.05.009. Epub 2011 May 11.
In this work, we proposed a novel biosensor to homogeneously detect concanavalin A (ConA) using pyrene-conjugated maltose assembled graphene based on fluorescence resonance energy transfer (FRET). Maltose-grafted-aminopyrene (Mal-Apy) was synthesized and characterized by mass spectra, UV-vis and fluorescence spectra. The Mal-Apy was further employed for fluorescence switch and ConA recognition. When Mal-Apy was self-assembled on the surface of graphene by means of π-stacking interaction, its fluorescence was adequately quenched because the graphene acted as a "nanoquencher" of the pyrene rings due to FRET. As a result, in the presence of ConA, competitive binding of ConA with glucose destroyed the π-stacking interaction between the pyrene and graphene, thereby causing the fluorescence recovery. This method was demonstrated the selective sensing of ConA, and the linear range is 2.0 × 10⁻² to 1.0 μM with the linear equation y=1.029x + 0.284 (R = 0.996). The limit of detection for ConA was low to 0.8 nM, and the detection of ConA could be performed in 5 min, indicating that this method could be used for fast, sensitive, and selective sensing of ConA. Such data suggests that the graphene FRET platform is a great potential application for protein-carbohydrate studies, and would be widely applied in drug screening, bimolecular recognition and disease diagnosis.
在这项工作中,我们提出了一种新的生物传感器,通过荧光共振能量转移(FRET)使用吡咯烯共轭麦芽糖组装的基于石墨烯均匀检测伴刀豆球蛋白 A(ConA)。通过质谱、紫外可见光谱和荧光光谱对麦芽糖接枝-氨基吡咯(Mal-Apy)进行了合成和表征。Mal-Apy 进一步用于荧光开关和 ConA 识别。当 Mal-Apy 通过 π-堆积相互作用自组装在石墨烯表面上时,由于 FRET,其荧光被充分猝灭,因为石墨烯作为吡咯环的“纳米猝灭剂”。结果,在 ConA 存在下,ConA 与葡萄糖的竞争性结合破坏了吡咯与石墨烯之间的 π-堆积相互作用,从而导致荧光恢复。该方法用于选择性检测 ConA,线性范围为 2.0×10⁻²至 1.0 μM,线性方程为 y=1.029x + 0.284(R = 0.996)。ConA 的检测限低至 0.8 nM,并且可以在 5 分钟内完成 ConA 的检测,表明该方法可用于快速、灵敏、选择性检测 ConA。这些数据表明,石墨烯 FRET 平台在蛋白质-碳水化合物研究中具有很大的应用潜力,并将广泛应用于药物筛选、双分子识别和疾病诊断。