中文摘要：丝状噬菌体是只感染细菌的病毒。本研究应用噬菌体展示技术鉴定高选择性的Cu（II）结合肽。经过五轮Cu(II)阳性筛选和多轮竞争金属离子(Al(III)、Co(II)、Fe(III)、Ni(II)和Zn (II)阴性筛选，富集噬菌体。通过酶联免疫吸附试验（ELISA）、扫描电镜（SEM）和X射线能谱（EDX）分析证实了最终噬菌体与Cu（II）的选择性结合。随机选取15个噬菌斑进行测序。选择出现频率最高、与Cu（II）亲和力最强的Cu-5肽（HGFANVA）进行进一步的Cu（II）检测和去除试验。电感耦合等离子体质谱（ICP-MS）证实了工程噬菌体具有很强的Cu（II）结合潜力。利用C端三个半胱氨酸单元合成了Cu-5肽，并基于AuNPs在Cu（II）离子作用下的聚集行为，建立了AuNP肽生物传感器系统。基于AuNP的Cu（II）传感器对Cu（II）具有选择性，LOD为91.15nM（约5.8 x 10(-3) mg/L），远低于世卫组织接受的1.3 mg/L的指导值。本研究提供了一种跨学科方法，将短肽应用于生物传感器研究的识别单元，具有用户友好、便捷、经济的特点。
外文摘要：Filamentous bacteriophages are viruses infecting only bacteria. In this study, phage display technique was applied to identify highly selective Cu(II) binding peptides. After five rounds of positive screening against Cu(II) and various rounds of negative screenings against competitive metal ions (Al(III), Co(II), Fe(III), Ni(II) and Zn (II)), bacteriophages were enriched. Selective Cu(II) binding of final phages was confirmed by Enzyme Linked Immunosorbent Assay (ELISA), Scanning Electron Microscopy (SEM) and Energy Dispersive X-ray spectroscopy (EDX) analyses. 15 phage plaques were randomly selected and sequenced. Cu-5 peptide (HGFANVA) with the highest frequency of occurrence and the strongest Cu(II) affinity was chosen for further Cu(II) detection and removal tests. Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) confirmed the strong Cu(II) binding potential of engineered viruses. Cu-5 peptides were synthetically synthesized with three Cysteine units at C-terminal and a AuNP-peptide biosensor system was developed based on aggregation behavior of AuNPs upon Cu (II) ion treatment. AuNP-based Cu(II) sensor was selective for Cu(II) and the LOD was 91.15 nM (ca. 5.8 x 10(-3) mg/L; 3 sigma/k, n = 5, R-2 = 0.992) for the case study which is considerably lower than the WHO's accepted guideline of 1.3 mg/L. This study provides an interdisciplinary approach to apply short peptides as recognition units for biosensor studies which are user friendly, not bulky and cost-effective.
外文关键词：Biosenso；Copper；Phage display；Genetic engineering；Filamentous phages
作者：Korkmaz, N；Hwang, C；Kessler, KK；Silina, YE；Muller, L；Park, J
作者单位：Europe Forsch Gesell MbH；Korea Univ；Univ Saarland；
期刊名称：TALANTA
期刊影响因子：5.339
出版年份：2021
出版刊次：232
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