Quantitative protein analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the selected reaction monitoring (SRM) mode was used to quantify matrix metalloprotease-9 (MMP-9; similar to 90 kDa) in bronchoalveolar lavage fluid (BALF) from patients having undergone lung transplantation. We developed an SRM assay for microfluidics-based nanoLC-MS/MS on a triple quadrupole mass spectrometer based on two signature peptides. Samples were prepared by chloroform-methanol precipitation followed by trypsin digestion in the presence of stable-isotope-labeled internal peptide standards. The method allows accurate quantification of MMP-9 in BALF with an LLOQ of 2.9 ng/mL and an LLOD of 0.25 ng/mL without the use of extensive fractionation or antibodies. (C) 2012 Elsevier B.V. All rights reserved.
|Tijdschrift||Journal of Chromatography A|
|Status||Published - 13-jul-2012|
|Evenement||37th International Symposium on High-Performance Liquid-Phase Separations and Related Techniques/18th NSEC/2nd DISEC - |
Duur: 8-okt-2011 → 11-okt-2011