Silica Colloidal Crystals for High Resolution MALDI-MS of Glycoproteins

DESCRIPTION provided by applicant The product is a slide coated with silica colloidal crystals to combine protein electrophoresis with matrix assisted laser desorption ionization mass spectrometry MALDI MS detection giving superior performance in both The Phase II proposal will be to develop an instrument for facile application of the slide in proteomics of intat glycoproteins This proposal is a collaboration between the Wirth group at Purdue who have developed new media for protein separations that can be coupled with MALDI MS and LI COR Biosciences whose core competency is the commercialization of instruments for bioanalysis Preliminary results using a modified silica colloidal crystalline surface as a MALDI MS substrate show a reduction in adduct formation and an increase in mass resolution Additionally the silica colloidal crystalline surface is a medium to separate proteins using isoelectric focusing Combining the ability to both separate and detect proteins using a single thin layer crystalline surface that enables high resolution protein separation and improved MALDI MS detection will expand the utility of using MALDI MS for identifying novel biomarkers based on post translational modifications The phase I specific aims are to optimize the silica crystal propertie to characterize the applicable molecular weight range to study the enhanced resolution and the reduction in adduct formation as a direct result of using this substrate and to combine protein separation techniques with MALDI MS detection on a single silica colloidal crystalline surface PUBLIC HEALTH RELEVANCE This project focuses on developing a glass slide coated with silica colloidal crystals that can serve as both an electrophoretic separation medium and an improved matrix assisted laser desorption ionization mass spectrometry MALDI MS target surface The crystalline surface integrates the methods of separating proteins by isoelectric focusing with MALDI MS detection producing a system that has both improved capabilities and improved peak resolution Advances in MALDI MS detection will aid in the analysis of post translational modifications and the discovery of novel biomarkers