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Quantification of Matrix Metalloprotease-9 in Bronchoalveolar Lavage Fluid by Selected Reaction Monitoring with Microfluidics Nano-Liquid-Chromatography–Mass Spectrometry

Prely et al., Journal of Chromatography (2012) - PMID: 22464002

Product(s) used in this publication:  SpikeTides™ - Proteomics Peptide Standards

Abstract:

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; ∼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.

Copyright © 2012 Elsevier B.V. All rights reserved.

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