DCP PhosProfile integrates phosphoproteomics with high-resolution quantitative mass spectrometry and bioinformatics to analyse the effects of drugs on the phosphoproteome and analysis of signal transduction pathways. PhosProfile allows for the unbiased identification and quantitation of regulated phosphorylation sites in the cellular proteome following exposure to small molecules that interact with cellular kinases or phosphatases.
Kinases form a major class of protein molecules that drive several signaling pathways involved in cell growth and differentiation. Aberrant activity of several kinases has now been attributed to the cause and progression of many cancers. For example, the EGFR family of receptor tyrosine kinases have been of great interest in recent years because of their involvement in several malignancies. Pharmacological inhibitors that are capable of binding and inhibiting these receptors are being extensively studied with a view of developing them into drugs to treat cancers that are caused or driven by these receptors. Her2(ERBB2) is one such receptor that has been associated with breast cancer. Though several Her2 inhibitors are now being tested in breast cancer patients with Her2 over-expression, the signaling intermediates and the effect of inhibitors on the pathways governed by Her2 are largely unknown. This issue has been addressed by performing a phosphoproteomics analysis of Her2 signaling using 3T3 cells with basal levels of Her2, 3T3s over-expressing Her2 and the over-expressing cells treated with pharmacological inhibitors.
Major applications of PhosProfile technology include:
- Target identification and validation: use PhosProfile to identify cellular kinase targets and to check if a hit from a phenotypic screen has a target involved in regulated protein phosphorylation
- Drug lead optimization
- Clinical candidate selection: use PhosProfile to determine if the kinase inhibitor clinical candidate is selective
- Drug mechanism and pathway identification: global analysis of cellular phosphorylation patterns upon drug treatment can provide information on drug mode of action and off target effects thus facilitating drug development.
Outline of Process for PhosProfile Analysis