(215a) Quick, Easy, Reproducible Protein Isoelectric Fractionation Using Protein Forest digital ProteomechipTM Technology

Fractionation is necessary to help decipher protein content in a complex sample. Protein Forest has developed a quick, reproducible and easy to use isoelectric fractionation technology: the digital ProteomeChip^TM (dPC^TM).

The dPC^TM consists of discrete pH gel features with pH intervals of 0.1 or smaller, which, when placed in an electric field for 30 min, will each trap and collect proteins simultaneously from a complex mixture according to the proteins' respective isoelectric points (pI). The resultant protein fractions can then be enzymatically-digested for subsequent MS identification, blotted to a membrane for antibody staining or transferred to traditional SDS-PAGE. The individual pH features allow researchers to reproducibly extract protein fractions from a complex mixture for detailed analysis, such as in biomarker discovery, protein-protein interaction research and clinical assay applications.

We have applied the dPC^TM technology to fractionate complex cell lysates and analyzed using LC/MS. By combining the dPC^TM technology with western analysis, we were able to perform pI separation on rat liver cell lysates and quickly determine the post-translational modification states of potential biomarker proteins. In this talk, we will discuss representative experimental results and other potential applications of the dPC^TM technology.