cIEF of glycoproteins in short nanoporous channels

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DESCRIPTION (provided by applicant): New technology to enable higher throughput in biomarker discovery is urgently needed. Medicine is rich in protein biomarker candidates for cancer, but poor in clinically validated biomarkers. The reason is that the methods for discovery are very low in throughput, usually giving false indications due to the statistics of small numbers Targeted searches of glycoforms are promising for the discovery of protein biomarkers for cancer. We propose to determine the feasibility of 3-mm channels packed with nonporous silica for rapid isoelectric focusing of the glycoforms of an immunoextracted protein from serum. These channels have a 96-well format for seamless integration into a fully automation analysis, from serum to detection, enabling thousands of serum samples per day to be characterized. Prostate specific antigen (PSA) is chosen as the model protein because there is published evidence that isoelectric focusing can resolve glycoforms with higher specificity for cancer, and there is a wealth of data and serum samples for this biomarker. We have established a collaboration with a clinical oncologist to obtain human serum samples from prostate cancer patients for testing the feasibility of this technology.