Integrated Sample Preparation and MALDI Mass Spectrometry on a Microfluidic Compact Disk
- 9 December 2003
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 76 (2) , 345-350
- https://doi.org/10.1021/ac030194b
Abstract
High-throughput microfluidic processing of protein digests integrated with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry on a compact disk (CD) is described. Centrifugal force moves liquid through multiple microstructures, each containing a 10-nL reversed-phase chromatography column. The CD enables parallel preparation of 96 samples with volumes ranging from one to several microliters. The peptides in the digests are concentrated, desalted, and subsequently eluted from the columns directly into MALDI target areas (200 × 400 μm) on the CD using a solvent containing the MALDI matrix. After crystallization, the CD is inserted into the MALDI instrument for peptide mass fingerprinting and database identification at a routine sensitivity down to the 200-amol level. Detection of proteolytic peptides down to the 50-amol level is demonstrated. The success rate of the CD technology in protein identification is about twice that of the C18 ZipTips and standard MALDI steel targets. The CDs are operated using robotics to transfer samples and reagents from microcontainers to the processing inlets on the disposable CD and spinning to control the movement of liquid through the microstructures.Keywords
This publication has 10 references indexed in Scilit:
- Thr94 in Bovine Myelin Basic Protein Is a Second Phosphorylation Site for 42-kDa Mitogen-Activated Protein Kinase (ERK2)Protein Journal, 2003
- Stability of Polycarbonate and Polystyrene Surfaces after Hydrophilization with High Intensity Oxygen RF PlasmaJournal of Colloid and Interface Science, 2002
- What place for polyacrylamide in proteomics?Trends in Biotechnology, 2001
- The Sequence of the Human GenomeScience, 2001
- Identification of foetal brain proteins by two‐dimensional gel electrophoresis and mass spectrometryEuropean Journal of Biochemistry, 2000
- Mass spectrometric identification of proteins and characterization of their post-translational modifications in proteome analysis.Analytical and Bioanalytical Chemistry, 2000
- Proteome mapping by two-dimensional polyacrylamide gel electrophoresis in combination with mass spectrometric protein sequence analysisPublished by Springer Nature ,2000
- Generating Electrospray from Microchip Devices Using Electroosmotic PumpingAnalytical Chemistry, 1997
- Matrix/sample interactions in ultraviolet laser‐desorption of proteinsRapid Communications in Mass Spectrometry, 1991
- Rational series of solvents for use in incremental gradient elutionAnalytical Chemistry, 1973