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Powering Proteomic Research with Automated Capillary Immunoassays

Wednesday, November 06, 2013 — Poster Session I

4:00 p.m. – 6:00 p.m.

FAES Academic Center (Upper-Level Terrace)

NCI

CANCER-2

Authors

  • J-Q Chen
  • M.A. Herrmann

Abstract

Capillary-based immunoassay systems (ProteinSimple NanoPro and Simple Western technologies) have been recently established in the Collaborative Protein Technology Resource (CPTR), a core facility in CCR/NCI, specializing in developing and implementing cutting edge protein-based technologies. These automated capillary immunoassay systems provide comprehensive and quantitative analysis to characterize proteins using nanogram amounts of material with good assay sensitivity and reproducibility. NanoPro is a capillary based iso-electric focusing (IEF) immunoassay system which employs high-resolution IEF followed by target specific immuno-probing to detect and quantify multiple protein phosphorylation isoforms using pan-reactive antibodies. NanoPro provides highly specific information on the activation status of signaling molecules that are not assessable by traditional western. The second system, Simple Western, is an automated western analysis performed in capillaries. Compared with conventional western, the Simple Western system offers easy operation, less sample consumption, highly quantitative data and excellent assay reproducibility. The potential of these technologies for clinically relevant biomarker development and therapeutic target identification is particularly exciting. Panels of over 100 assays covering key signaling pathways have been developed in our core and applied to the profiling of signaling events in response to targeted therapies both in vitro cell culture and in vivo mouse xenograft and human clinical specimens.

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