期刊
SENSORS
卷 20, 期 18, 页码 -出版社
MDPI
DOI: 10.3390/s20185275
关键词
magnetic graphene oxide (MGO); proteases; in situ peptide synthesis; fluorescence resonance energy transfer (FRET); biological assays
资金
- Ministry of Trade, Industry and Energy, Korea [10077599]
- Basic Science Research Program through the National Research Foundation of Korea (NRF) - Ministry of Education [2019R1F1A1062208, 2017R1A6A1A03015642]
- Korea Evaluation Institute of Industrial Technology (KEIT) [10077599] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
- National Research Foundation of Korea [2019R1F1A1062208] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
Graphene oxide (GO)/peptide complexes as a promising disease biomarker analysis platform have been used to detect proteolytic activity by observing the turn-on signal of the quenched fluorescence upon the release of peptide fragments. However, the purification steps are often cumbersome during surface modification of nano-/micro-sized GO. In addition, it is still challenging to incorporate the specific peptides into GO with proper orientation using conventional immobilization methods based on pre-synthesized peptides. Here, we demonstrate a robust magnetic GO (MGO) fluorescence resonance energy transfer (FRET) platform based on in situ sequence-specific peptide synthesis of MGO. The magnetization of GO was achieved by co-precipitation of an iron precursor solution. Magnetic purification/isolation enabled efficient incorporation of amino-polyethylene glycol spacers and subsequent solid-phase peptide synthesis of MGO to ensure the oriented immobilization of the peptide, which was evaluated by mass spectrometry after photocleavage. The FRET peptide MGO responded to proteases such as trypsin, thrombin, and beta-secretase in a concentration-dependent manner. Particularly, beta-secretase, as an important Alzheimer's disease marker, was assayed down to 0.125 ng/mL. Overall, the MGO platform is applicable to the detection of other proteases by using various peptide substrates, with a potential to be used in an automated synthesis system operating in a high throughput configuration.
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