Department of Chemistry, Hacettepe University, Ankara, Turkey.
Faculty of Pharmacy, Ege University, İzmir, Turkey.
Methods Mol Biol. 2022;2393:37-56. doi: 10.1007/978-1-0716-1803-5_3.
Nucleic acid aptamers are an emerging class of artificial ligands and have recently gained attention in several areas. Here we report the design of a surface plasmon resonance (SPR) aptasensor for highly sensitive and selective sensing of human activated protein C (APC). First, DNA aptamer (DNA-Apt) specific for APC is complexed with N-methacryloyl-L-cysteine (MAC) monomer. Then, 2-hydroxyethyl methacrylate (HEMA) and cyanamide are mixed with the DNA-Apt/MAC complex. The SPR aptasensor is characterized by atomic force microscopy, ellipsometry, and contact angle measurements. Selectivity of SPR aptasensor is carried out in the presence of myoglobin (Myb), hemoglobin (Hb), and bovine serum albumin (BSA). Limit of detection (LOD) and limit of quantification (LOQ) values are 1.5 ng mL and 5.2 ng mL, respectively. DNA-Apt SPR aptasensor performance for APC detection is also examined in artificial plasma.
核酸适体是一类新兴的人工配体，最近在多个领域引起了关注。在这里，我们报告了一种用于高灵敏度和选择性检测人激活蛋白 C (APC) 的表面等离子体共振 (SPR) 适体传感器的设计。首先，与 APC 特异性结合的 DNA 适体 (DNA-Apt) 与 N-丙烯酰-L-半胱氨酸 (MAC) 单体复合。然后，将 2-羟乙基甲基丙烯酸酯 (HEMA) 和氰胺与 DNA-Apt/MAC 复合物混合。SPR 适体传感器的特性通过原子力显微镜、椭圆测量法和接触角测量法进行表征。在肌红蛋白 (Myb)、血红蛋白 (Hb) 和牛血清白蛋白 (BSA) 存在的情况下进行 SPR 适体传感器的选择性检测。检测限 (LOD) 和定量限 (LOQ) 值分别为 1.5ng mL 和 5.2ng mL。还在人工血浆中检查了 APC 检测用 DNA-Apt SPR 适体传感器的性能。
