Difference Gel Electrophoresis of Phosphoproteome

Research and Development labs (R&D) in academia, pharmaceutical, and biotech companies often assess the phosphorylation of a protein or classes of proteins. Currently, large-scale phosphorylation analysis is used to understand how signaling pathways work and how they may be deregulated in disease states. Even though thousands of protein phosphorylation sites can be identified, researchers may only be interested in a small sample of proteins for a signaling pathway. Hence, these large scale analysis techniques are time-consuming and cost-prohibitive. The applicability of gel-based proteomic strategy in phosphoproteomics has been largely limited by the lack of technologies for specific and quantitative detection of phosphoproteins in gels.

To resolve these issues, researchers at Purdue University have developed a novel gel-based process, called Differential Gel Electrophoresis of Phosphoproteome (DiGEP). This process uses metal ions, such as Ti4+, Zr4+, Fe3+, and Ga3+, which are attracted to phosphate groups and attach fluorophores on the phosphoproteins. This ultimately helps visualize phosphorylation changes in different samples on the same gel. DiGEP analyses are advantageous because they enable the visualization of phosphoproteins on a single gel and select only relevant proteins for in-gel digestion and mass spectrometric analysis. It is highly specific, selective, and quantitative and can be used routinely in labs for quantitative phosphorylation measurement, in vitro kinase assay, kinase and phosphatase activity assay, kinase/phosphatase inhibitor screening, and detection of in vivo phosphorylation. Not only will this technology help a general biological laboratory to effectively measure changes in protein phosphorylation, but will also help pharmaceutical and biotech industries to develop effective diagnostic and therapeutic agents relating to kinases.

Advantages:

-Visualization of phosphoproteins

-Selection of proteins for spectrometric analysis

-Effective and quantitative measures

Potential Applications:

-R&D labs

-Disease research

TRL: 4

Intellectual Property:

Keywords: DiGEP, Differential Gel Electrophoresis of Phosphoproteome, protein phosphorylation analysis, phosphoproteomics, gel-based proteomic strategy, quantitative phosphorylation measurement, in vitro kinase assay, kinase phosphatase activity assay, kinase/phosphatase inhibitor screening, in vivo phosphorylation detection