中文摘要：本文研制了一种涂有多肽的电化学生物传感器，可以检测流感病毒（H5N1）包膜中的血凝素（HA）蛋白。利用噬菌体展示技术，鉴定了对HA具有特异性结合能力的高亲和力肽展示噬菌体颗粒（GHPHYNNPSLQL），并用酶联免疫吸附试验（ELISAs）进行了评价。根据功能研究确定的分子结构，化学合成了6种新设计的多肽，并用原子力显微镜（AFM）、扫描电子显微镜（SEM）、等温滴定量热法（ITC）和表面等离子体共振光谱（SPR）对其与HA蛋白的结合作用进行了表征。方波伏安法（SWV）显示电流变化与HA蛋白浓度呈线性关系。据此，选择IFA-BP1肽作为潜在的分子受体，并证实检测限（LOD）为3.26nm。此外，电化学传感器成功地检测到人血浆加标样品中的HA；在31.2-500nm的浓度范围和82.3-111.5%的浓度范围内，使用该生物传感器具有良好的重复性和回收率。该传感器系统测得的人血浆样品中HA蛋白的水平与使用商业ELISA法结果相当，并具有制造成本低、分析速度快和样品体积较低等优点，可用于流感病毒抗原的快速有效检测，这点尤为重要。
外文摘要：In this study, we developed an electrochemical biosensor coated with a peptide that can detect the protein hemagglutinin (HA) present in the envelope of the influenza virus (H5N1). Using phage display, high-affinity peptide-displaying phage particles (GHPHYNNPSLQL) with specific binding abilities for HA were identified, which were evaluated using enzyme-linked immunosorbent assays (ELISAs). Based on the molecular structure identified by the functional studies, six newly designed peptides were chemically synthesized, and their binding interactions with HA proteins were characterized by atomic force microscopy (AFM), scanning electron microscopy (SEM), isothermal titration calorimetry (ITC), and surface plasmon resonance spectroscopy (SPR). Square wave voltammetry (SWV) revealed the presence of a linear relationship between the change in current and concentration of HA protein. From these observations, we selected IFA BP1 peptide as a potential molecular receptor and confirmed that the limit of detection (LOD) was found to be 3.26 nM. In addition, our electrochemical sensor successfully detected HA spiked in human plasma samples. Furthermore, good results for reproducibility and recovery using this biosensor were observed within a range of 31.2-500 nM concentrations and 82.3-111.5%, respectively. Importantly, the levels of HA proteins in the samples of human plasma measured using this sensor system corresponded well with those determined using a commercial ELISA method and also offered advantages such as low cost of fabrication, faster analysis, and use of lower sample volumes, suggesting that it could be applied for rapid and effective detection of the influenza virus antigen.
外文关键词：Influenza virus；Hemagglutinin；Affinity peptide；Molecular receptor；Peptide sensor
作者：Kim, JH；Cho, CH；Shin, JH；Hyun, MS；Hwang, E；Park, TJ；Park, JP
作者单位：Chung Ang Univ；Natl NanoFab Ctr NNFC；Korea Basic Sci Inst
期刊名称：SENSORS AND ACTUATORS B-CHEMICAL
期刊影响因子：7.1
出版年份：2021
出版刊次：343
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