Discovery Proteomics

Partners: CNG, Digilab GmbH, Roche, OGT

Genome-wide protein profiling, or ‘proteomic’ technology platforms are used by the MolPAGE partners to obtain an understanding of global patterns of protein content and activity at a given time in tissues and biofluid samples and to determine how protein expression profiles change in response to disease.  WP5 are developing a range of protein and peptide analysis platforms which they will apply to the samples obtained by WP1 to identify protein and peptide markers associated with intermediate phenotypes of T2D and CVD.

Biomarker discovery scans will be performed using three analysis platforms incorporating an initial protein fractionation step to simplify subsequent biomarker identification.  For 2D gel mass fingerprinting, pre-fractionated protein extracts are separated on 2D gels. After staining spots are excised, digested with trypsin, and analysed by MALDI mass spectrometry. Differential peptide display (DPD) relies of the fractionation of extracts by HPLC followed by analysis using mass spectrometry (ESI and MALDI). In clinical proteomics the fractionation of the proteins is carried out using the different affinity of coated magnetic beads, and fraction extracts subjected to mass spectrometry (MALDI).  A goal of WP5 is to establish an automated platform for clinprot proteomics with a view to preparing this technology for application on an epidemiological scale.