Mass spectrometry-based in vitro kinome activity assay
This technology is a new generation, high-throughput assay to quantitatively assess the activity of the kinome in a variety of samples. Cocktails of synthetic peptides are combined with a sample of interest and phosphorylated by kinases in the sample. The degree of phosphorylation is then measured by ultra-high resolution mass-spectrometry.
The methods optimize the speed and accuracy of high-throughput mass spectrometry analysis. For example, the peptides in the cocktail are enriched with stable isotopes to enhance the ionization efficiency. The end result is an assay capable of measuring the activity of kinases from single cells. Combined with cell sorting, this assay could provide a powerful means to analyze subsets of cells from tissue samples.
The lab is currently working to expand the peptide cocktail and is seeking collaborators to apply the technology to disease diagnosis and prognosis.
Intellectual Property Status: Patent(s) Pending
Protein kinases are important cellular proteins involved in a wide range of signaling networks. Dysregulation of protein kinase signaling pathways is a hallmark of many human diseases, including cancer. Measuring normal or abnormal kinase activity is important to understanding the biology of these diseases. Although the regulation and functions of many individual kinases are well understood, high-throughput methods to assess the activity of the full complement of protein kinases, the kinome, remain scarce.
The screen includes peptides associated with certain cancers, cardiac conditions and inflammatory conditions, making the assay useful for diagnosis and prognosis, as well as for patient stratification for clinical trials. This assay could also be used for discovery research to profile kinase activity in disease tissues, discover new targets for kinase inhibitors, and establish relationships between the activity of particular kinases and clinical outcomes.