Combination of MALDI-TOF mass spectrometry with immobilized enzyme microreactor for peptide mapping

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has been combined with immobilized enzyme microreactor for the rapid, sensitive, and accurate tryptic mapping of protein and polypeptides. The technique utilizes the trypsin microreactor by immobilized enzyme on the glycidyl methacrylate (GMA)-modified cellulose membrane. The membrane micro-reactor was used for the tryptic mapping of cytochrome C and the results were compared with those obtained by using free trypsin. A significant increase in the overall sensitivity of the process was observed using the membrane microreactor, as well as the elimination of background signals due to the autolysis of the trypsin. Further, membrane microreactor digestions were found to be rapid and convenient.