Trypsin immobilization on an ethylenediamine-based monolithic minidisk for rapid on-line peptide mass fingerprinting studies.
The aim of this work was to develop a trypsin-based micro-immobilized enzyme reactor prepared on a monolithic ethylenediamine BIA Separations CIM (convective interaction media) minidisk. The micro-immobilized enzyme reactor (IMER) was integrated in a liquid chromatography system hyphenated to electrospray ionization tandem mass spectrometry to carry out on-line protein digestion and identification. The performance of this IMER was compared with that obtained using a previously developed bioreactor prepared on a conventional CIM ethylenediamine disk and with that of the commercially available Poroszyme immobilized trypsin cartridge. In this work, we showed how different proteins were identified with good recoveries using a digestion time of 10 min only.
Nicoli
R
R
Laboratory of Pharmaceutical Analytical Chemistry, School of Pharmaceutical Sciences, University of Geneva, University of Lausanne, 20 Boulevard d'Yvoy, 1211 Geneva 4, Switzerland.
Rudaz
S
S
Stella
C
C
Veuthey
J-L
JL
D002853:Chromatography, Liquid; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005029:Ethylenediamines / Q000737:chemistry*; D010449:Peptide Mapping; D011506:Proteins / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000737:chemistry*
D004800:Enzymes, Immobilized; D005029:Ethylenediamines; D011506:Proteins; C031234:ethylenediamine; D014357:Trypsin