Multiplexed protein and kinase activity detection .. (MPKADAIC)
Multiplexed protein and kinase activity detection assays in complex media using impedance/capacitance
Start date: Nov 1, 2011,
End date: Oct 31, 2013
"The detection and quantification of protein biomarkers in biological samples lies central to proteomics, drug design, disease prognosis and therapeutic development. The generation of viable protein microarrays is, though, challenging. The first protein microarrays were built onantibodies. Unfortunately antibodies do not function well in the microarray format, because typically only a small fraction (20%) specifically recognizes the target protein. Current antibody based optical assays are commonly based on sandwich assays in which antigen binding to the immobilised antibody is detected through the use of a secondary, labeled, antibody. Though sensitive, this method is laborious and often requires a specifically-labelled secondary antibody for every antigen of interest.Labelling protocols are potentially perturbative, can also be time consuming and may lead to high background signals. Alternative protein receptive molecules are thus of considerable interest. In recent years, the host group has developed, with a team in Leeds, optical and electrical assays based on the use of peptide aptamers (highly specific protein receptors built into the surface of robust scaffold proteins). These can be immobilized with controlled surface orientation on a variety of surfaces. The aim of this proposal is to utilize this experience in developing highly sensitive electrical protein assays using capacitance and impedance (AC, DC, Faradaic and Non Faradaic). Through appropriate surface chemical methods such assays will be operable in complex fluid such as cell lysates and blood. Being electrical they are also readily multiplexed at comparatively low cost, enabling simultaneous detection of multiple targets. The linear range potentiallyaccessible within such arrays is considerable, as is the potential clinical benefit."
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