Upconversion Fluorescence Resonance Energy Transfer Based Biosensor for Ultrasensitive Detection of Matrix Metalloproteinase-2 in Blood

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• 作者：Yuhui Wang ; Pei Shen ; Chunya Li ; Yanying Wang ; Zhihong Liu
• 刊名：Analytical Chemistry
• 出版年：2012
• 出版时间：February 7, 2012
• 年：2012
• 卷：84
• 期：3
• 页码：1466-1473
• 全文大小：360K
• 年卷期：v.84,no.3(February 7, 2012)
• ISSN：1520-6882

文摘

Matrix metalloproteinase-2 (MMP-2) is a very important biomarker in blood. Presently, sensitive and selective determination of MMP-2 directly in blood samples is still a challenging job because of the high complexity of the sample matrix. In this work, we reported a new homogeneous biosensor for MMP-2 based on fluorescence resonance energy transfer (FRET) from upconversion phosphors (UCPs) to carbon nanoparticles (CNPs). A polypeptide chain (NH₂-GHHYYGPLGVRGC-COOH) comprising both the specific MMP-2 substrate domain (PLGVR) and a 蟺-rich motif (HHYY) was designed and linked to the surface of UCPs at the C terminus. The FRET process was initiated by the 蟺鈥撓€ interaction between the peptide and CNPs, which thus quenched the fluorescence of the donor. Upon the cleavage of the substrate by the protease at the amide bond between Gly and Val, the donor was separated from the acceptor while the 蟺-rich motif stayed on the acceptor. As a result, the fluorescence of the donor was restored. The fluorescence recovery was found to be proportional to the concentration of MMP-2 within the range from 10鈥?00 pg/mL in an aqueous solution. The quantification limit of this sensor was at least 1 order of magnitude lower than that of other reported assays for MMP-2. The sensor was used to determine the MMP-2 level directly in human plasma and whole blood samples with satisfactory results obtained. Owing to the hypersensitivity of the method, clinical samples of only less than 1 渭L were needed for accurate quantification, which can be meaningful in MMP-2-related clinical and bioanalytical applications.