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Proceedings Paper

The use of time-resolved fluorescence in gel-based proteomics for improved biomarker discovery
Author(s): AnnSofi Sandberg; Volker Buschmann; Peter Kapusta; Rainer Erdmann; Åsa M. Wheelock
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Paper Abstract

This paper describes a new platform for quantitative intact proteomics, entitled Cumulative Time-resolved Emission 2-Dimensional Gel Electrophoresis (CuTEDGE). The CuTEDGE technology utilizes differences in fluorescent lifetimes to subtract the confounding background fluorescence during in-gel detection and quantification of proteins, resulting in a drastic improvement in both sensitivity and dynamic range compared to existing technology. The platform is primarily designed for image acquisition in 2-dimensional gel electrophoresis (2-DE), but is also applicable to 1-dimensional gel electrophoresis (1-DE), and proteins electroblotted to membranes. In a set of proof-of-principle measurements, we have evaluated the performance of the novel technology using the MicroTime 100 instrument (PicoQuant GmbH) in conjunction with the CyDye minimal labeling fluorochromes (GE Healthcare, Uppsala, Sweden) to perform differential gel electrophoresis (DIGE) analyses. The results indicate that the CuTEDGE technology provides an improvement in the dynamic range and sensitivity of detection of 3 orders of magnitude as compared to current state-of-the-art image acquisition instrumentation available for 2-DE (Typhoon 9410, GE Healthcare). Given the potential dynamic range of 7-8 orders of magnitude and sensitivities in the attomol range, the described invention represents a technological leap in detection of low abundance cellular proteins, which is desperately needed in the field of biomarker discovery.

Paper Details

Date Published: 24 February 2010
PDF: 5 pages
Proc. SPIE 7568, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VIII, 756811 (24 February 2010); doi: 10.1117/12.842571
Show Author Affiliations
AnnSofi Sandberg, Karolinska Institutet (Sweden)
Volker Buschmann, PicoQuant GmbH (Germany)
Peter Kapusta, PicoQuant GmbH (Germany)
Rainer Erdmann, PicoQuant GmbH (Germany)
Åsa M. Wheelock, Karolinska Institutet (Sweden)


Published in SPIE Proceedings Vol. 7568:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VIII
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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