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  • P-ISSN 1225-0163
  • E-ISSN 2288-8985

Acoustic technology-assisted rapid proteolysis for high-throughput proteome analysis

Analytical Science and Technology / Analytical Science and Technology, (P)1225-0163; (E)2288-8985
2011, v.24 no.6, pp.510-518
https://doi.org/10.5806/AST.2011.24.6.510






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Abstract

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.

keywords
LC/MS/MS, protein digestion, acoustic technology


Reference

1

1. R. Aebersold and D. R. Goodlett, Chem. Rev., 101, 269-295 (2001).

2

2. R. Aebersold and M. Mann, Nature, 422, 198-207 (2003).

3

3. T. J Griffin, D. R. Goodlett, R. Aebersold and C. Opin, Biotech., 12, 607-612 (2001).

4

4. A.-J. Moulay A. and Y. J. Xu, J. Zhejiang Univ. Science B, 7(6), 411 (2006).

5

5. J. M. Gilmore and M. P. Washburn, J. Proteomics, 73(11), 2078-2091 (2010).

6

6. J. A. Loo, C.G. Edmonds and R. D. Smith, Anal. Chem., 63, 2488-2499 (1991).

7

7. J. R. III Yates, J. Mass Spectrometry, 33, 1-19 (1998).

8

8. J. P. Chang, D. E. Kiehl and A. Kennington, Rapid Commun. Mass Spectrom., 11, 1266-1270 (1997).

9

9. G. W. Slysz and D. C. Schriemer, Anal. Chem., 77(6), 1572-1579 (2005).

10

10. I. M. Lazar, R. S. Ramsey and J. M. Ramsey, Anal. Chem., 73(8), 1733-1739 (2001).

11

11. W. Sun, S. Gao, L. Wang, Y. Chen, S. Wu, X. Wang, D. Zheng and Y. Gao, Mol. Cel. Proteomics, 5, 769-776 (2006).

12

12. G. Yao, C. Deng, X. Zhang and P. Yang, Angew. Chem. Int. Ed, 49, 8185 (2010).

13

13. B. E. Slentz, N. A. Penner and F. E. Regnier, J.Chromatogr. A, 984(1), 97-107 (2003).

14

14. E. Bonneil, M. Mercier and K. C. Waldron, Anal. Chim. Acta, 404, 29-45 (2000).

15

15. L. J. Jin, J. Ferrance, J. C. Sanders and J. P. Landers, Lab. Chip, 3, 11-18 (2003).

16

16. K. Yamada, T. Nakasone, R. Nagano and M. Hirata, J. Appl. Polym. Sci., 89, 3574-3581 (2003).

17

17. K. Sakai-Kato, M. Kato and T. Toyo'oka, Anal. Chem., 74, 2943 (2002).

18

18. J. Gao, J. Xu, L. E. Locascio and C.S. Lee, Anal. Chem., 73, 2648-2655 (2001).

19

19. D. S. Peterson, T. Rohr, F.Svec and J. M. J. Frechet, Anal. Chem., 74, 4081-4088 (2002).

20

20. G. W. Slysz and D. C. Schriemer, Rapid Commun Mass Spectrom., 17, 1044-1050 (2003).

21

21. http://www.covarisinc.com/how_it_works.html

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