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Journal of Biomedicine and Biotechnology
Volume 2011, Article ID 213643, 7 pages
Research Article

The Utilization of Triton X-100 for Enhanced Two-Dimensional Liquid-Phase Proteomics

1Department of Microbiology, Chungbuk National University, 410 Sungbong-Ro, Heungduk-Gu, Cheongju 361-763, Republic of Korea
2Department of Chemical Engineering, Chonbuk National University, 664-14, 1-Ga, Duckjin-Dong, Duckjin-Gu, Jeonju 561-156, Republic of Korea
3School of Natural System, College of Science and Engineering, Kanazawa University, Kakuma-Machi, Kanazawa, Ishikawa 920-1192, Japan

Received 16 May 2011; Revised 8 August 2011; Accepted 9 August 2011

Academic Editor: J. R. Botella

Copyright © 2011 Mina Kim et al. 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.


One of the main challenges in proteomics lies in obtaining a high level of reproducible fractionation of the protein samples. Automated two-dimensional liquid phase fractionation (PF2D) system manufactured by Beckman Coulter provides a process well suited for proteome studies. However, the protein recovery efficiency of such system is low when a protocol recommended by the manufacturer is used for metaproteome profiling of environmental sample. In search of an alternative method that can overcome existing limitations, this study replaced manufacturer's buffers with Triton X-100 during the PF2D evaluation of Escherichia coli K12. Three different Triton X-100 concentrations—0.1%, 0.15%, and 0.2%—were used for the first-dimension protein profiling. As the first-dimension result was at its best in the presence of 0.15% Triton X-100, second-dimension protein fractionation was performed using 0.15% Triton X-100 and the standard buffers. When 0.15% Triton X-100 was used, protein recovery increased as much as tenfold. The elution reliability of 0.15% Triton X-100 determined with ribonuclease A, insulin, α-lactalbumin, trypsin inhibitor, and cholecystokinin (CCK) affirmed Triton X-100 at 15% can outperform the standard buffers without having adverse effects on samples. This novel use of 0.15% Triton X-100 for PF2D can lead to greater research possibilities in the field of proteomics.