Rapid and Ultra - Sensitive Determination of Enzyme Activities Using Surface Enhanced Resonance Raman Scattering  

Oleh:

Moore, Barry D. ; Stevenson, Lorna ; Watt, Alan ; Flitsch, Sabine ; Turner, Nicolas J. ; Cassidy, Chris ; Graham, Duncan

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Nature Biotechnology: The Science and Business of Biotechnology vol. 22 no. 9 (Sep. 2004), page 1133-1138.
Topik: scattering; ultra sensitive determination; enzyme; surface enhance; resonance; raman scattering

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: NN9.1 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

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Isi artikel

Measurement of enzyme activity and selectivity at in vivo concentrations is highly desirable in a range of field including diagnostics, functional proteomics and directed evolution. Here we demonstrate how surface enhanced resonance Raman scattering (SERRS), measured using silver nanoparticles, can be used to detect the activity of hydrolases at ultra low levels. This approach was made possible by designing masked enzyme substrates that are initially completely undetected by SERRS. Turnover of the substrate by the enzyme leads to the release of a surface targeting dye, and intense SERRS signals proportional to enzyme actvity are generated. The method was used to rapidly screeen the relative activities and enantioselectivities of fourteen enzymes including examples of lipases, esterases and proteases. In the current format the sensitivity of the technique is sufficient to detect 500 enzyme molecules, which offers the potential to detect multiple enzyme activities simulataneously and at levels found within single cells.

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