Bertok Tomas, Lorencova Lenka, Hroncekova Stefania, Gajdosova Veronika, Jane Eduard, Hires Michal, Kasak Peter, Kaman Ondrej, Sokol Roman, Bella Vladimir, Eckstein Anita Andicsova, Mosnacek Jaroslav, Vikartovska Alica, Tkac Jan
Institute of Chemistry, Slovak Academy of Sciences, Dubravska cesta 9, Bratislava 845 38, Slovak Republic.
Glycanostics Ltd., Dubravska cesta 9, Bratislava 845 38, Slovak Republic.
MethodsX. 2019 Sep 11;6:1999-2012. doi: 10.1016/j.mex.2019.08.017. eCollection 2019.
The article describes preparation, characterization and further modification of hybrid magnetic particles (Au nanoshells with a magnetic core (MPs@silica@Au)) by zwitterionic molecules bearing diazonium functional groups. Such hybrid magnetic particles modified by zwitterionic molecules exhibit the following features: •Responsiveness towards external magnetic field applicable for various enrichment strategies due to magnetic core;•Golden outer layer exhibiting free surface plasmons could be used for grafting of zwitterionic molecules diazonium functionality;•Zwitterionic interface on such particles provides resistivity towards non-specific protein binding; and at the same time such interface was applied for immobilization of antibodies against prostate specific antigen (PSA) applied for selective enrichment of PSA from serum samples with subsequent electrochemical assays. The approach presented here using hybrid magnetic particles can be easily applied for immobilization of antibodies using a highly robust surface patterning protocols by formation of a self-assembled monolayer with delivery of functional groups on the outer surface of magnetic particles. Hybrid magnetic particles with immobilized antibodies are applied for highly efficient and quick separation of protein of interest PSA from complex sample. Finally, hybrid magnetic particles with "fished-out" protein molecules could be incubated with lectins to form a sandwich configuration for glycoprofiling of PSA.
本文描述了通过带有重氮官能团的两性离子分子对杂化磁性颗粒(具有磁性核心的金纳米壳(MPs@二氧化硅@金))进行制备、表征及进一步修饰的过程。经两性离子分子修饰的此类杂化磁性颗粒具有以下特性:•由于磁性核心,对外部磁场有响应,适用于各种富集策略;•具有自由表面等离子体激元的金色外层可用于两性离子分子重氮官能团的接枝;•此类颗粒上的两性离子界面可抵抗非特异性蛋白质结合;同时该界面用于固定抗前列腺特异性抗原(PSA)的抗体,以从血清样本中选择性富集PSA,随后进行电化学分析。本文介绍的使用杂化磁性颗粒的方法可通过在磁性颗粒外表面形成具有官能团递送功能的自组装单分子层,利用高度稳健的表面图案化方案轻松用于抗体的固定。固定有抗体的杂化磁性颗粒用于从复杂样品中高效快速分离目标蛋白质PSA。最后,带有“钓出”蛋白质分子的杂化磁性颗粒可与凝集素孵育,形成夹心结构用于PSA的糖谱分析。