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http://hdl.handle.net/2445/149617
Title: | On-line protein digestion by immobilized enzyme microreactor capillary electrophoresis-mass spectrometry |
Author: | Villegas, Lorena Peró Gascón, Roger Benavente Moreno, Fernando J. (Julián) Barbosa Torralbo, José Sanz Nebot, María Victoria |
Keywords: | Electroforesi capil·lar Enzims Espectrometria de masses Capillary electrophoresis Enzymes Mass spectrometry |
Issue Date: | 10-Feb-2019 |
Publisher: | Elsevier B.V. |
Abstract: | In this study, we present the use of microreactors packed with immobilized trypsin particles for the rapid and efficient bottom-up analysis of proteins by on-line immobilized enzyme microreactor capillary electrophoresis mass spectrometry (IMER-CE-MS). The results obtained digesting β-lactoglogulin (β-LG) off-line with free trypsin in solution and with immobilized trypsin particles were taken as a reference for the optimization of the on-line protein digestion. Under the optimized conditions, on-line digestion, separation and characterization of the protein digests were possible in less than 30 min. The limit of detection for complete sequence coverage was around 10 µg mL-1 (~500 µM) of β-LG, the repeatability was comparable to the off-line digestion methods and the microreactor could be reused until thirty times. The good performance of IMER-CE-MS was also demonstrated for several other proteins as α-casein (α-CSN), β-casein (β-CSN), and κ-casein (κ-CSN), as well as for a complex protein mixture (an Escherichia coli whole cell lysate). |
Note: | Versió postprint del document publicat a: https://doi.org/10.1016/j.talanta.2019.02.039 |
It is part of: | Talanta, 2019, vol. 199, p. 116-123 |
URI: | http://hdl.handle.net/2445/149617 |
Related resource: | https://doi.org/10.1016/j.talanta.2019.02.039 |
ISSN: | 0039-9140 |
Appears in Collections: | Articles publicats en revistes (Enginyeria Química i Química Analítica) |
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