Quaglio Deborah, Mangiardi Laura, Venditti Giulia, Del Plato Cristina, Polli Francesca, Ghirga Francesca, Favero Gabriele, Pierini Marco, Botta Bruno, Mazzei Franco
Department of Chemistry and Technology of Drugs, Department of Excellence 2018-2022, Sapienza University of Rome, P.le Aldo Moro 5, 00185, Rome, Italy.
Center For Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Viale Regina Elena 291, 00161, Rome, Italy.
Chemistry. 2020 Jul 8;26(38):8400-8406. doi: 10.1002/chem.202000989. Epub 2020 Jun 25.
One of the main problems in the development of immunosensors is to overcome the complexity of binding antibodies to the sensor surface. Most immobilizing methods lead to a random orientation of antibodies with a lower binding site density and immunoaffinity. In order to control the orientation of antibody immobilization, several resorc[4]arene derivatives were designed and synthesized. After the spectroscopic characterization of resorc[4]arene self-assembled monolayers (SAMs) onto gold films, the surface coverage and the orientation of insulin antibody (Ab-Ins) were assessed by a surface plasmon resonance (SPR) technique and compared with a random immobilization method. Experimental results combined with theoretical studies confirmed the dipole-dipole interaction as an important factor in antibody orientation and demonstrated the importance of the upper rim functionalization of resorcarenes. Accordingly, resorcarene 5 showed a major binding force towards Ab-Ins thanks to the H-bond interactions with the amine protein groups. Based on these findings, the resorcarene-based immunosensor is a powerful system with improved sensitivity providing new insight into sensor development.
免疫传感器开发中的主要问题之一是克服将抗体结合到传感器表面的复杂性。大多数固定方法会导致抗体随机取向，结合位点密度和免疫亲和力较低。为了控制抗体固定的取向，设计并合成了几种间苯二酚[4]芳烃衍生物。在对金膜上的间苯二酚[4]芳烃自组装单分子层（SAMs）进行光谱表征后，通过表面等离子体共振（SPR）技术评估了胰岛素抗体（Ab-Ins）的表面覆盖率和取向，并与随机固定方法进行了比较。实验结果与理论研究相结合，证实了偶极-偶极相互作用是抗体取向的重要因素，并证明了间苯二酚上缘功能化的重要性。因此，由于与胺蛋白基团的氢键相互作用，间苯二酚5对Ab-Ins表现出主要的结合力。基于这些发现，基于间苯二酚的免疫传感器是一个强大的系统，具有更高的灵敏度，为传感器开发提供了新的见解。
