A new application of microwave technology to proteomics

Abstract

Two-dimensional electrophoresis (2-DE) combined with mass spectrometry has significantly improved the possibilities of large-scale identification of proteins. However, 2-DE is limited by its inability to speed up the in-gel digestion process. We have developed a new approach to speed up the protein identification process utilizing microwave technology. Proteins excised from gels are subjected to in-gel digestion with endoprotease trypsin by microwave irradiation, which rapidly produces peptide fragments. The peptide fragments were further analyzed by matrix-assisted laser desorption/ionization technique for protein identification. The efficacy of this technique for protein mapping was demonstrated by the mass spectral analyses of the peptide fragmentation of several proteins, including lysozyme, albumin, conalbumin, and ribonuclease A. The method reduced the required time for in-gel digestion of proteins from 16 hours to as little as five minutes. This new application of microwave technology to protein identification will be an important advancement in biotechnology and proteome research.

Microwave‐assisted proteomics

“…It was observed that a 15-min enzymatic incubation time was sufficient for high peptide recovery and protein coverage. This concept was further validated by Juan et al (2005), who demonstrated optimum conditions for in-gel microwave-assisted tryptic digestions at 378C for 5 min.…”

Digestion completeness of microwave-assisted and conventional trypsin-catalyzed reactions

“…This approach exploits unique microwave properties to accelerate the enzymatic digestion. In recent years, microwaveassisted proteolytic digestion procedures, including tryptic [6][7][8] and acid-mediated proteolysis [9][10][11][12], have enabled samples to be prepared more rapidly for bottom-up analysis.…”

Microwave Assisted Reactions in Organic Chemistry: A Review of Recent Advances

“…They have observed that the interaction between protein or enzyme and 2.45 GHz electromagnetic field is responsible for these differences and macroscopic change in temperature has no role over them (Lill, 2009;Zhao, 2010;Lill et al, 2007;Sandoval et al, 2007;. Besides the solution (Lill, 2009;Zhao, 2010;Lill et al, 2007;Sandoval et al, 2007;Sandoval et al, 2008), in-gel tryptic digests also demonstrated enhanced digest efficiency/sequence coverage (Sun et al, 2005;Juan et al, 2005). If we load digests with magnetic elements to enhance the microwave absorptivity of the irradiated solutions, similar improvements in terms of digest efficiency/sequence coverage could be observed (Y.…”