金纳米粒子(AuNP)在生物技术中起着至关重要的作用,医学,和诊断由于其独特的光学性质。它们与抗体结合,抗原,蛋白质,或核酸能够实现精确靶向并增强生物传感能力。功能化的AuNP,然而,可能会降低稳定性,导致聚集或功能丧失,特别是在复杂的生物环境中。此外,它们可以显示与非预期目标的非特异性结合,损害测定特异性。在这项工作中,柠檬酸盐稳定和二氧化硅涂层的AuNP(GNP和SiGNP,分别)已使用N,基于N-二甲基丙烯酰胺的共聚物增加其稳定性并使其能够被生物分子官能化。修饰后的AuNP稳定性已通过包括分光光度表征在内的技术组合进行评估,纳米粒子跟踪分析,透射电子显微镜和功能微阵列测试。确定了两种不同的共聚物,以提供稳定的AuNP涂层,同时通过点击化学反应进行进一步修饰。由于聚合物中存在叠氮基团。按照这个实验设计,合成了用ssDNA和链霉亲和素修饰的AuNP,并成功用于生物测定。总之,已开发出一种用于AuNP的功能化方案,该方案易于修改,通常需要单一的步骤和短的孵化时间。获得的功能化AuNP提供了相当大的灵活性,因为功能化方案可以个性化以匹配多个测定的要求。
Gold nanoparticles (AuNPs) play a vital role in biotechnology, medicine, and diagnostics due to their unique optical properties. Their conjugation with antibodies, antigens, proteins, or nucleic acids enables precise targeting and enhances biosensing capabilities. Functionalized AuNPs, however, may experience reduced stability, leading to aggregation or loss of functionality, especially in complex biological environments. Additionally, they can show non-specific binding to unintended targets, impairing assay specificity. Within this work, citrate-stabilized and silica-coated AuNPs (GNPs and SiGNPs, respectively) have been coated using N,N-dimethylacrylamide-based copolymers to increase their stability and enable their functionalization with biomolecules. AuNP stability after modification has been assessed by a combination of techniques including spectrophotometric characterization, nanoparticle tracking analysis, transmission electron microscopy and functional microarray tests. Two different copolymers were identified to provide a stable coating of AuNPs while enabling further modification through click chemistry reactions, due to the presence of azide groups in the polymers. Following this experimental design, AuNPs decorated with ssDNA and
streptavidin were synthesized and successfully used in a biological assay. In conclusion, a functionalization scheme for AuNPs has been developed that offers ease of modification, often requiring single steps and short incubation time. The obtained functionalized AuNPs offer considerable flexibility, as the functionalization protocol can be personalized to match requirements of multiple assays.