BioPRIA, Australian Pulp and Paper Institute (APPI), Department of Chemical Engineering, Monash University, Clayton, VIC 3800, Australia.
J Colloid Interface Sci. 2013 Nov 1;409:59-65. doi: 10.1016/j.jcis.2013.07.051. Epub 2013 Aug 2.
Bioactive papers are usually challenged by four major limitations: sensitivity, selectivity, simplicity and strength (4S). Gold nanoparticles (AuNPs) treated paper has previously been demonstrated as a Surface Enhanced Raman Scattering (SERS) active substrate, capable of addressing the 4S issues. In this study, AuNPs on paper substrate were functionalized by a series of biomolecules to develop a generic SERS platform for antibody-antigen detection. The functionalization steps were performed by taking advantage of the high affinity association between Streptomyces avidinii-derived protein, streptavidin, and biotin. Streptavidin was firstly bound onto the AuNPs treated paper using biotinylated-thiol. Subsequently, desired biotinylated-antibody was bound onto the streptavidin. SERS spectra of each functionalization step were obtained to ensure specific adsorption of the bio-molecules. The binding interaction of the antibody with its specific antigen was detected using SERS. Shifts of Raman band associated with α-helix and β-sheet structures indicated structural modification of the antibody upon interaction with its antigen. Predominant tryptophan and tyrosine residue bands were also detected, confirming the presence of antigen. Reproducible spectral features were quantified as AuNP papers were subjected to different concentrations of antigen; the spectra intensity increased as a function of the antigen concentration. The retention of AuNPs on paper remained constant after all the consecutive washing and functionalization steps. The feasibility of AuNPs paper as a low-cost and generic SERS platform for bio-diagnostic applications was demonstrated.
灵敏度、选择性、简单性和强度(4S)。金纳米粒子(AuNPs)处理过的纸张以前已被证明是表面增强拉曼散射(SERS)的活性衬底,能够解决 4S 问题。在这项研究中,AuNPs 在纸衬底上通过一系列生物分子进行功能化,以开发用于抗体-抗原检测的通用 SERS 平台。功能化步骤是利用链霉菌属avidinii 衍生的蛋白(链霉亲和素)和生物素之间的高亲和力结合来完成的。首先,使用生物素化硫醇将链霉亲和素结合到 AuNPs 处理过的纸张上。随后,将所需的生物素化抗体结合到链霉亲和素上。获得每个功能化步骤的 SERS 光谱,以确保生物分子的特异性吸附。使用 SERS 检测抗体与其特异性抗原的结合相互作用。与α-螺旋和β-折叠结构相关的拉曼带的位移表明抗体与抗原相互作用时结构发生了修饰。还检测到主要的色氨酸和酪氨酸残基带,证实了抗原的存在。随着抗原浓度的增加,AuNP 纸的光谱强度增加,作为定量的重现性光谱特征。在所有连续的洗涤和功能化步骤之后,AuNPs 在纸张上的保留率保持不变。AuNPs 纸作为用于生物诊断应用的低成本和通用 SERS 平台的可行性得到了证明。