G03019: An investigation into the use of MudPIT as a tool for the separation and quantification of proteins for GM crop safety assessments
Thursday 9 February 2006
This research project aims to assess the potential of multidimensional protein identification technology (MudPIT) as quantitative procedure to ascertain protein perturbations arising from genetic modifications.
Background
Despite advances in gel-based approaches to proteomic analysis, the technology remains unsuitable for the rigorous safety assessment of GM plants. Limiting factors include the restrictive type of proteins that can be analysed (i.e. hydrophobic proteins are poorly detected), inadequate dynamic range and inaccurate quantitation. MudPIT is an attractive alternative. The procedure has been shown to detect low abundance and hydrophobic proteins and quantitative methodologies for MudPIT have been developed.
Research Approach
The first stage of the project will optimise 'front-end' procedures in order to maximise proteome coverage by MudPIT. Both non chemical (e.g.Stable Isotope labelled Standards) and chemical (e.g. Isobaric Tag for Relative and Absolute Quantification (iTRAQ) approaches to quantification will be evaluated using complementary MS Techniques. The unique collection of GM material and plants available at RHUL will be used to fully evaluate and validate the procedure.