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Application of MesoScale Technology for the Measurement of Multiple Biomarkers to Explore Molecular and Cellular Processess.

Reference	BB/F010850/1
Principal Investigator / Supervisor	Professor Stephen Ward
Co-Investigators / Co-Supervisors	Professor Geoffrey Holman, Dr Amanda MacKenzie, Professor David Tosh, Dr Andrew Watts, Professor Melanie Welham, Ms Sue Wonnacott
Institution	University of Bath
Department	Pharmacy and Pharmacology
Funding type	Research
Value (£)	93,388
Status	Completed
Type	Research Grant
Start date	17/04/2008
End date	16/04/2009
Duration	12 months

Abstract

The Mesoscale Discovery (MSD) SECTOR Imager (SI) 6000 is a unique protein profiling system that offers a sensitive, adaptable platform for sandwich immunoassays in microplates with 1-10 carbon electrode arrays per well. This format enables multiplexed assays where several analytes (in multi-well plate format) can be rapidly measured. Ruthenium (Ru)-labeled antibodies are used in combination with the co-reactant tripropylamine contained. An electrochemical reaction is initiated when current is applied to the plates and the light signal emitted from Ru-label bound to capture electrodes is measured and quantified. The main features provided by the instrument are: - the ability to accurately quantify activation of specific signals; - it facilitates detection of multiple signals simultaneously. - the system allows for more rapid analysis, - provides platform for developing our own custom made assays for future applications. This instrument will be applied in a range of related project applications that address different molecular and cellular events essential to the Healthy Organism BBSRC key area: 1. Contribution of PI3K-dependent signaling events to T Lymphocyte function. 2. Analysis of macrophage P2X7 receptors and their significance to inflammatory signalling pathways . 3. Inflammatory signalling in response to influenza infection and drug treatment. 4. Analysis of molecular signals regulating ES Cell self-renewal and differentiation 5 Development of a 3-D Mouse pancreatic culture system with high throughput screening for anti-diabetic drugs. 6. Investigation of signalling steps involved in insulin action and insulin resistance in heart muscle cells. 7. Nicotinic receptor signalling through ERK/MAPK pathways.

Summary

Analysis of molecular and cellular events is crucial in modern life sciences research, for a better understanding of (i) fundamental biological processes, (ii) the changes that characterise associated disease states, and (iii) the development of therapeutics. This research proposal brings together seven research areas, spanning the regulation of the immune and nervous systems, haemopoiesis, tissue development and regeneration and cell metabolism that share a common theme of wanting to utilise sensitive, quantitative and high throughput state-of-the-art instrumentation to analyse a range of cellular mechanisms. This instrument is the Mesoscale Discovery (MSD) SECTOR Imager (SI) 6000 which offers a sensitive, adaptable platform with which to measure several analytes in cell or cell-derived samples. The technology relies on the use of chemically tagged antibodies that recognize the analytes of choice. When bound to their target, the Abs emit light upon electrochemical stimulation which is measured and quantified. The main features provided by the instrument are: - the ability to accurately quantify activation of specific signals; - it facilitates detection of multiple signals simultaneously. - in situations where cell samples may be limiting, information on several different signalling molecules can be generated simultaneously. - the system allows for more rapid analysis, - provides platform for developing our own custom made assays for future applications.

Committee	Closed Committee - Biochemistry & Cell Biology (BCB)
Research Topics	Pharmaceuticals
Research Priority	X – Research Priority information not available
Research Initiative	Research Equipment Initiative 2007 (RE7) [2007]
Funding Scheme	X – not Funded via a specific Funding Scheme

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