Soares Juliana Coatrini, Shimizu Flavio Makoto, Soares Andrey Coatrini, Caseli Luciano, Ferreira Jacqueline, Oliveira Osvaldo N
†São Carlos Institute of Physics, University of São Paulo, 369, 13560-970 São Carlos, São Paulo, Brazil.
‡Institute of Environmental, Chemical and Pharmaceutical Sciences, Federal University of Sao Paulo, 09972-970 Diadema, São Paulo, Brazil.
ACS Appl Mater Interfaces. 2015 Jun 10;7(22):11833-41. doi: 10.1021/acsami.5b03761. Epub 2015 May 29.
The need for early detection of various diseases, including breast cancer, has motivated research into nanomaterials that can be assembled in organized films which serve as biosensors. Owing to the variety of possible materials and film architectures, procedures are required to design optimized biosensors. In this study, we combine surface-specific methods to monitor the assembly of antibodies on nanostructured films with two distinct architectures. In the first, a layer of the antibody type mouse anti-HER2 (clone tab250) was immobilized on a self-assembled monolayer (SAM) of 11-mercaptoundecanoic acid modified with N-hydroxysuccinimide (NHS) and 1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide (EDC). In the second approach, a SAM of cysteamine was coated with a biotin/spreptavidin bilayer on which a layer of biotinylated antibody type MSx2HUp185/her biotin was adsorbed. The rougher, less passivating coating with cysteamine determined from cyclic voltammetry and scanning electron microscopy led to biosensors that are more sensitive to detect the breast cancer ERBB2 (HER2) biomarker in impedance spectroscopy measurements. This higher distinguishing ability of the cysteamine-containing film architecture was proven with information visualization methods to treat the impedance data. Polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) confirmed that biosensing resulted from the antibody-ERBB2 antigen affinity.
对包括乳腺癌在内的各种疾病进行早期检测的需求,推动了对可组装成用作生物传感器的有序薄膜的纳米材料的研究。由于可能的材料和薄膜结构种类繁多,因此需要一些程序来设计优化的生物传感器。在本研究中,我们将表面特异性方法结合起来,以监测抗体在具有两种不同结构的纳米结构薄膜上的组装情况。在第一种方法中,将一层小鼠抗HER2抗体(克隆tab250)固定在经N-羟基琥珀酰亚胺(NHS)和1-乙基-3-(3-(二甲基氨基)丙基)碳二亚胺(EDC)修饰的11-巯基十一烷酸自组装单分子层(SAM)上。在第二种方法中,用生物素/链霉亲和素双层涂覆半胱胺的SAM,在其上吸附一层生物素化的MSx2HUp185/her生物素抗体类型。通过循环伏安法和扫描电子显微镜确定的半胱胺涂层更粗糙、钝化作用更小,从而使生物传感器在阻抗谱测量中对检测乳腺癌ERBB2(HER2)生物标志物更敏感。通过信息可视化方法处理阻抗数据,证明了含半胱胺薄膜结构具有更高的区分能力。偏振调制红外反射吸收光谱(PM-IRRAS)证实生物传感是由抗体-ERBB2抗原亲和力引起的。