Capillary Immunoassays: New Promise for Proteomics Research in the Lab and in the Clinic

Thursday, October 11, 2012 — NIH Core Poster Session
10:00 a.m. – Noon	Building 10 South Lobby	NCI	CORE-2

Authors

• JQ Chen
• M.A. Herrmann

Abstract

Two capillary based immunoassay systems have recently been established in the Collaborative Protein Technology Resource, a core facility in CCR/NCI. First, the Nano(fluidic)Pro(teomics) technology, is a capillary-based isoelectric-focusing(IEF) immunoassay, which employs high-resolution IEF separation followed by target-specific immunoprobing to detect and quantify multiple protein phosphorylation isoforms that are not readily assessed by traditional Western blotting. This capillary-based system allows protein analysis in extremely small and precious samples, such as stem cells, primary cells, fine needle aspirates, and other patient specimens. NanoPro assays of many key signaling pathways, (e.g. MAPK, AKT, PKC, STAT and Caspase pathways) have been developed in our core, and successfully applied to the profiling of signaling events in response to pathway-targeted treatment in both preclinical and clinical samples, creating new opportunities for pharmacodynamic biomarker assessment in clinical studies. The second system, Simple Western system, is a fully automated, gel free, blot free Western system. Simple Western system greatly minimizes the variability caused by manual processes in traditional Western. Highly quantitative data and excellent reproducibility have been obtained with the system. A method for absolute quantification and direct comparison of protein isoform amount in cell lysates was developed to provide an approach of correlating protein quantity with function.

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