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Proteomes 2015, 3(4), 454-466; doi:10.3390/proteomes3040454

Bead Based Proteome Enrichment Enhances Features of the Protein Elution Plate (PEP) for Functional Proteomic Profiling

1
Array Bridge, Inc., St. Louis, MO 63108, USA
2
Biotech Support Group LLC, Monmouth Junction, NJ 08852, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Jacek R. Wisniewski
Received: 8 October 2015 / Revised: 19 November 2015 / Accepted: 30 November 2015 / Published: 8 December 2015
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Abstract

A novel functional proteomics technology called PEP(Protein Elution Plate) was developed to separate complex proteomes from natural sources and analyze protein functions systematically. The technology takes advantage of the powerful resolution of two-dimensional gel electrophoresis (2-D Gels). The modification of electrophoretic conditions in combination with a high-resolution protein elution plate supports the recovery of functionally active proteins. As 2DE(2-Dimensional Electrophoresis) resolution can be limited by protein load, we investigated the use of bead based enrichment technologies, called AlbuVoid™ and KinaSorb™ to determine their effect on the proteomic features which can be generated from the PEP platform. Using a variety of substrates and enzyme activity assays, we report on the benefits of combining bead based enrichment to improve the signal report and the features generated for Hexokinase, Protein Kinase, Protease, and Alkaline Phosphatase activities. As a result, the PEP technology allows systematic analysis of large enzyme families and can build a comprehensive picture of protein function from a complex proteome, providing biological insights that could otherwise not be observed if only protein abundances were analyzed. View Full-Text
Keywords: functional proteomics; 2-D gel; PEP; biomarkers; protein enrichment; cancer functional proteomics; 2-D gel; PEP; biomarkers; protein enrichment; cancer
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Wang, X.; Davies, M.; Roy, S.; Kuruc, M. Bead Based Proteome Enrichment Enhances Features of the Protein Elution Plate (PEP) for Functional Proteomic Profiling. Proteomes 2015, 3, 454-466.

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