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KMID : 0385520110240060510
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Analytical Science & Technology
2011 Volume.24 No. 6 p.510 ~ p.518
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Acoustic technology-assisted rapid proteolysis for high-throughput proteome analysis
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Kim Bo-ra
Trang Tran Huyen
Han Na-Young
Park Jong-Moon
Yu Ung-Sik
Lee Hoo-keun
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Abstract
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Recent developments and improvements of multiple technological elements including mass spectrometry (MS) instrument, multi-dimensional chromatographic separation, and software tools processing MS data resulted in benefits of large scale proteomics analysis. However, its throughput is limited by the speed and reproducibility of the protein digestion process. In this study, we demonstrated a new method for rapid proteolytic digestion of proteins using acoustic technology. Tryptic digests of BSA prepared at various conditions by super acoustic for optimization time and intensity were analyzed by LC-MS/MS showed higher sequence coverage in compared with traditional 16 hrs digestion method. The method was applied successfully for complex proteins of a breast cancer cells at 30 min of digestion at intensity 2. This new application reduces timeconsuming of sample preparation with better efficiency, even with large amount of proteins, and increases highthroughput process in sample preparation state.
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KEYWORD
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LC/MS/MS, protein digestion, acoustic technology,
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