Sosnowska Marta, Łęga Tomasz, Olszewski Marcin, Gromadzka Beata
Department of Analysis and Chemical Synthesis, Institute of Biotechnology and Molecular Medicine, Kampinoska 25., 80-180, Gdańsk, Poland.
Nano Expo Sp z.o.o, Kładki 24, 80-822, Gdańsk, Poland.
Microb Cell Fact. 2025 Jan 8;24(1):11. doi: 10.1186/s12934-024-02553-4.
Ecotoxicology is essential for the evaluation and comprehension of the effects of emergency pollutants (EP) such as heavy metal ions on the natural environment. EPs pose a substantial threat to the health of humans and the proper functioning of the global ecosystem. The primary concern is the exposure of humans and animals to heavy metal ions through contaminated water. The presence of heavy metal ions in drinking water ought to be monitored in accordance with World Health Organization regulations. Among the numerous harmful metal ions, copper ions are responsible for a variety of human diseases.
This study investigates the application of phage display as a screening method for heavy metal toxicological targets, with copper served as the main focus. To identify a variety of Cu-binding M13 phage clones with unique peptides and to assess their affinity for metal ions, the study utilized Escherichia coli as a factories producing recombinant bacteriophages, modified biopanning procedure and an ELISA assay. The research highlights the increasing importance of phage display as a screening tool in ecotoxicology. We synthesized and modified the selected peptide to enable the rapid optical detection of Cu(II) ions in aqueous solutions. By incorporating the dansyl group into a designated peptide sequence, we implemented fluorescence detection assays for real-time measurements. The Cu- binding peptide's efficacy was confirmed through spectroscopic measurements, which allowed for real-time detection with rapid response times with high selectivity.
The phage display technique was successfully applied to develop the fluorescent peptide-based chemosensor that exhibited high selectivity and sensitivity for Cu.
生态毒理学对于评估和理解重金属离子等应急污染物（EP）对自然环境的影响至关重要。应急污染物对人类健康和全球生态系统的正常功能构成重大威胁。主要担忧的是人类和动物通过受污染的水接触重金属离子。饮用水中重金属离子的存在应按照世界卫生组织的规定进行监测。在众多有害金属离子中，铜离子会引发多种人类疾病。
本研究以铜为主要研究对象，探讨噬菌体展示作为重金属毒理学靶点筛选方法的应用。为了鉴定出具有独特肽段的多种结合铜的M13噬菌体克隆，并评估它们对金属离子的亲和力，该研究利用大肠杆菌作为生产重组噬菌体的工厂，采用改良的生物淘选程序和酶联免疫吸附测定法。该研究突出了噬菌体展示作为生态毒理学筛选工具日益重要的地位。我们合成并修饰了所选肽段，以便能够快速光学检测水溶液中的铜离子（Cu(II)）。通过将丹磺酰基团引入指定的肽序列中，我们实施了荧光检测测定法以进行实时测量。通过光谱测量证实了铜结合肽的功效，其能够以快速的响应时间进行高选择性的实时检测。
噬菌体展示技术成功应用于开发对铜具有高选择性和灵敏度的基于荧光肽的化学传感器。
