Department of Chemistry, Nanchang University, Nanchang 330031, China.
Analyst. 2014 Jan 21;139(2):416-24. doi: 10.1039/c3an01818k.
In recent years, great efforts have focused on the exploration and fabrication of protein nanoconjugates due to potential applications in many fields including bioanalytical science, biosensors, biocatalysis, biofuel cells and bio-based nanodevices. An important aspect of our understanding of protein nanoconjugates is to quantitatively understand how proteins interact with nanomaterials. In this report, human serum albumin (HSA) and citrate-coated silver nanoparticles (AgNPs) are selected as a case study of protein-nanomaterial interactions. UV-visible spectroscopy together with multivariate curve resolution by alternating least squares (MCR-ALS) algorithm is first exploited for the detailed study of AgNPs-HSA interactions. Introduction of the chemometrics tool allows extracting the kinetic profiles, spectra and distribution diagrams of two major absorbing pure species (AgNPs and AgNPs-HSA conjugate). These resolved profiles are then analysed to give the thermodynamic, kinetic and structural information of HSA binding to AgNPs. Transmission electron microscopy, circular dichroism spectroscopy and Fourier transform infrared spectroscopy are used to further characterize the complex system. Moreover, a sensitive spectroscopic biosensor for HSA is fabricated with the MCR-ALS resolved concentration of absorbing pure species. It is found that the linear range for the HSA nanosensor was from 1.9 nM to 45.0 nM with a detection limit of 0.9 nM. It is believed that the proposed method will play an important role in the fabrication and optimization of a robust nanobiosensor or cross-reactive sensors array for the detection and identification of biocomponents.
近年来，由于在生物分析科学、生物传感器、生物催化、生物燃料电池和基于生物的纳米器件等多个领域的潜在应用，人们致力于探索和制备蛋白质纳米缀合物。了解蛋白质纳米缀合物的一个重要方面是定量了解蛋白质如何与纳米材料相互作用。在本报告中，人血清白蛋白（HSA）和柠檬酸包覆的银纳米颗粒（AgNPs）被选为蛋白质-纳米材料相互作用的案例研究。首先利用紫外-可见光谱结合交替最小二乘法（MCR-ALS）算法的多元曲线分辨来详细研究 AgNPs-HSA 相互作用。引入化学计量学工具允许提取两个主要吸收纯物质（AgNPs 和 AgNPs-HSA 缀合物）的动力学谱、光谱和分布图。然后分析这些分辨出的谱图，以给出 HSA 与 AgNPs 结合的热力学、动力学和结构信息。透射电子显微镜、圆二色光谱和傅里叶变换红外光谱用于进一步表征复杂体系。此外，利用 MCR-ALS 分辨出的吸收纯物质浓度制备了用于检测 HSA 的灵敏光谱生物传感器。结果发现，HSA 纳米传感器的线性范围为 1.9 nM 至 45.0 nM，检测限为 0.9 nM。相信该方法将在用于检测和鉴定生物成分的稳健纳米生物传感器或交叉反应传感器阵列的制造和优化中发挥重要作用。
