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Gold surface supported spherical liposome-gold nano-particle nano-composite for label free DNA sensing

机译:金表面负载球形脂质体-金纳米颗粒纳米复合材料,用于无标记DNA传感

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摘要

Immobilization of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) liposome-gold nano-particle (DOPE-AuNP) nano-composite covalently on 3-mercaptopropionic acid (MPA) on gold surface is demonstrated for the first time for electrochemical label free DNA sensing. Spherical nature of the DOPE on the MPA monolayer is confirmed by the appearance of sigmoidal voltammetric profile, characteristic behavior of linear diffusion, for the MPA-DOPE in presence of [Fe(CN)_6]~(3-/4-) and [Ru(NH_3)_6]~(3+) redox probes. The DOPE liposome vesicle fusion is prevented by electroless deposition of AuNP on the hydrophilic amine head groups of the DOPE. Immobilization of single stranded DNA (ssDNA) is made via simple gold-thiol linkage for DNA hybridization sensing in the presence of [Fe(CN)_6]~(3-/4-). The sensor discriminates the hybridized (complementary target hybridized), un-hybridized (non-complementary target hybridized) and single base mismatch target hybridized surfaces sensitively and selectively without signal amplification. The lowest target DNA concentration detected is 0.1×10~(-12)M. Cyclic voltammetry (CV), electrochemical impedance (EIS), differential pulse voltammetry (DPV) and quartz crystal microbalance (QCM) techniques are used for DNA sensing on DOPE-AuNP nano-composite. Transmission Electron Microscopy (TEM), Fourier Transform Infrared Spectroscopy (FTIR), Atomic Force Microscopy (AFM), Dynamic Light Scattering (DLS) and Ultraviolet-Visible (UV) spectroscopic techniques are used to understand the interactions between the DOPE, AuNP and ssDNA. The results indicate the presence of an intact and well defined spherical DOPE-AuNP nano-composite on the gold surface. The method could be applied for fabrication of the surface based liposome-AuNP-DNA composite for cell transfection studies at reduced reagents and costs.
机译:首次证明了在金表面的3-巯基丙酸(MPA)上共价固定有1,2-二油酰基-sn-甘油-3-磷酸乙醇胺(DOPE)脂质体-金纳米颗粒(DOPE-AuNP)纳米复合物。用于无电化学标记的DNA感测。在[Fe(CN)_6]〜(3- / 4-)和[[]存在下,MPA-DOPE的S形伏安轮廓,线性扩散的特征行为证实了DOPE在MPA单层上的球形性质。 Ru(NH_3)_6]〜(3+)氧化还原探针。通过在DOPE的亲水胺头基上化学沉积AuNP来防止DOPE脂质体囊泡融合。在[Fe(CN)_6]〜(3- / 4-)存在下,通过简单的金硫醇键固定DNA杂交,从而固定单链DNA(ssDNA)。传感器可以灵敏而有选择地区分杂交(互补靶杂交),非杂交(非互补靶杂交)和单碱基错配靶杂交表面,而无需信号放大。检测到的最低目标DNA浓度为0.1×10〜(-12)M。循环伏安法(CV),电化学阻抗(EIS),差分脉冲伏安法(DPV)和石英晶体微量天平(QCM)技术用于DOPE-AuNP纳米复合材料的DNA感测。使用透射电子显微镜(TEM),傅立叶变换红外光谱(FTIR),原子力显微镜(AFM),动态光散射(DLS)和紫外可见(UV)光谱技术来了解DOPE,AuNP和ssDNA之间的相互作用。结果表明在金表面上存在完整且轮廓分明的球形DOPE-AuNP纳米复合材料。该方法可用于制备基于表面的脂质体-AuNP-DNA复合材料,用于细胞转染研究,且试剂减少,成本降低。

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