期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 114, 期 34, 页码 9056-9061出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1700317114
关键词
surface-enhanced Raman spectroscopy; SERS; protein detection; nanoparticle; ELISA
资金
- Institute for Collaborative Biotechnologies from the US Army Research Office [W911NF-09-D-0001]
- Defense Advanced Research Projects Agency [N66001-14-2-4055]
- US Army Medical Research and Material Command, Defense Medical Research and Army Research Office [W911NF-10-2-0114]
- Omnis Global Technologies
We present a sensitive and quantitative protein detection assay that can efficiently distinguish between specific and nonspecific target binding. Our technique combines dual affinity reagents with surface-enhanced Raman spectroscopy (SERS) and chemometric analysis. We link one Raman reporter-tagged affinity reagent to gold nanoparticles and another to a gold film, such that protein-binding events create a hot spot with strong SERS spectra from both Raman reporter molecules. Any signal generated in this context is indicative of recognition by both affinity labels, whereas signals generated by nonspecific binding lack one or the other label, enabling us to efficiently distinguish true from false positives. We show that the number of hot spots per unit area of our substrate offers a quantitative measure of analyte concentration and demonstrate that this dual-label, SERS-linked aptasensor assay can sensitively and selectively detect human alpha-thrombin in 1% human serum with a limit of detection of 86 pM.
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