Abstract
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometric (MALDI-TOF MS) analysis of proteins in salt-containing solution was performed for the first time using porous anodic alumina (PAA) membrane as sample support. The resulting spectral quality of proteins under standard sample preparation conditions was superior to that of normal metal sample stages. Analysis of phosphate-doped protein solutions indicated that porous anodic alumina membranes as a target yielded better results than a metallic target for salt-containing solutions. Because of the biocompatibility of the PAA, proteins can be adsorbed on the PAA and thus a washing process can be introduced to remove the salts from the PAA target before MS analysis. This desalting step significantly enhanced spectral quality, and better signal-to-noise ratios were obtained. The present technique is promising for proteomics research.
Article PDF
Avoid common mistakes on your manuscript.
References
Karas, M.; Hillenkamp, F. Laser Desorption Ionization of Proteins with Molecular Masses Exceeding 10,000 Daltons. Anal. Chem. 1988, 60, 2299–2301.
Tanaka, K.; Waki, H.; Ido, Y.; Akita, S.; Yoshida, Y.; Yoshida, T. Protein and Polymer Analyses up to m/z 100,000 by Laser Ionization Time-of-flight Mass Spectrometry. Rapid Commun. Mass Spectrom. 1988, 2, 151–153.
Laugesen, S.; Roepstorff, P. Combination of Two Matrices Results in Improved Performance of MALDI MS for Peptide Mass Mapping and Protein Analysis. J. Am. Soc. Mass Spectrom. 2003, 14, 992–1002.
Fenselau, C. MALDI MS and Strategies for Protein Analysis. Anal. Chem. 1997, 69, 661A-665A.
Hillenkamp, F.; Karas, M.; Beavis, R. C.; Chait, B. T. Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry of Biopolymers. Anal. Chem. 1991, 63, 1193A-1203A.
Chou, J. Z.; Kreek, M. J.; Chait, B. T. Matrix-Assisted Laser Desorption Mass Spectrometry of Biotransformation Products of Dynorphin A in Vitro. J. Am. Soc. Mass Spectrom. 1994, 5, 10–16.
Warren, M. E.; Brockman, A. H.; Orlando, R. On-Probe Solid-Phase Extraction/MALDI-MS Using Ion-Pairing Interactions for the Cleanup of Peptides and Proteins. Anal. Chem. 1998, 70, 3757–3761.
Brockman, A. H.; Dodd, B. S.; Orlando, R. A. Desalting Approach for MALDI-MS Using On-Probe Hydrophobic Self-Assembled Monolayers. Anal. Chem. 1997, 69, 4716–4720.
Winston, R. L.; Fitzgerald, M. C. Concentration and Desalting of Protein Samples for Mass Spectrometry Analysis. Anal. Biochem. 1998, 262, 83–85.
Xu, Y. D.; Watson, J. T.; Bruening, M. L. Patterned Monolayer/Polymer Films for Analysis of Dilute or Salt-Contaminated Protein Samples by MALDI-MS. Anal. Chem. 2003, 75, 85–190.
Zhang, L.; Orlando, R. Solid-Phase Extraction/MALDI-MS: Extended Ion-Pairing Surfaces for the On-Target Cleanup of Protein Samples. Anal. Chem. 1999, 71, 4753–4757.
Mock, K. K.; Sutton, C. W.; Cottrell, J. S. Sample Immobilization Protocols for Matrix-Assisted Laser Desorption Mass Spectrometry. Rapid Commun. Mass Spectrom. 1992, 6, 233–238.
Xu, Y. D.; Bruening, M. L.; Watson, J. T. Nonspecific, On-Probe Cleanup Methods for MALDI-MS Samples. Mass Spectrom. Rev. 2003, 22, 429–440.
Oleschuk, R. D.; McComb, M. E.; Chow, A.; Ens, W.; Standing, K. G.; Perreault, H.; Marois, Y.; King, M. Characterization of Plasma Proteins Adsorbed onto Biomaterials by MALDI-TOFMS. Biomaterials 2000, 21, 1701–1710.
Vestling, M. M.; Fenselau, C. Poly(vinylidene difluoride) Membranes as the Interface Between Laser Desorption Mass Spectrometry, Gel Electrophoresis, and in Situ Proteolysis. Anal. Chem. 1994, 66, 471–477.
Kurien, B. T.; Matsumoto, H.; Scofield, R. H. Purification of Tryptic Peptides for Mass Spectrometry Using Polyvinylidene Fluoride Membrane. Ind. J. Biochem. Biophys. 2001, 38, 274–276.
Blackledge, J. A.; Alexander, A. J. Polyethylene Membrane as a Sample Support for Direct Matrix-Assisted Laser Desorption/Ionization Mass Spectrometric Analysis of High Mass Proteins. Anal. Chem. 1995, 67, 843–848.
Zaluzec, E. J.; Gage, D. A.; Allison, J.; Watson, J. T. Direct Matrix-Assisted Laser Desorption Ionization Mass Spectrometric Analysis of Proteins Immobilized on Nylon-Based Membranes. J. Am. Soc. Mass Specctrom. 1994, 5, 203–237.
McComb, M. E.; Oleschuk, R. D.; Manley, D. M.; Donald, L.; Chow, A.; O’Neil, J. D. J.; Ens, W.; Standing, K. G.; Perreault, H. Use of a Nonporous Polyurethane Membrane as a Sample Support for Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry of Peptides and Proteins. Rapid Commun. Mass Spectrom. 1997, 11, 1716–1722.
Thompson, G. E.; Fumeaux, R. C.; Wood, G. C.; Richardson, J. A.; Goode, J. S. Nucleation and Growth of Porous Anodic Films on Aluminium. Nature 1978, 272, 433–435.
Masuda, H.; Fukuda, K. Ordered Metal Nanohole Arrays Made by a Two-Step Replication of Honeycomb Structures of Alumina. Science 1995, 268, 1466–1468.
Yuan, J. H.; He, F. Y.; Sun, D. C.; Xia, X. H. A. Simple Method for Preparation of Through-Hole Porous Anodic Alumina Membrane. Chem. Mater. 2004, 16, 1841–1844.
Martin, C. R. Nanomaterials: A Membrane-Based Synthetic Approach. Science 1994, 266, 1961–1966.
Chio, J.; Sauer, G.; Nielsch, K.; Wehspohn, R. B.; Gosele, U. Hexagonally Arranged Monodisperse Silver Nanowires with Adjustable Diameter and High Aspect Ratio. Chem. Mater. 2003, 15, 776–779.
Reiss, B. D.; Freeman, R. G.; Walton, I. D.; Norton, S. M.; Smith, P. C.; Stonas, W. G.; Keating, C. D.; Natan, M. J. Electrochemical Synthesis and Optical Readout of Striped Metal Rods with Submicron Features. J. Eletroanal. Chem. 2002, 522, 95–103.
Yuan, J. H.; Wang, K.; Xia, X. H. Highly Ordered Platinum Nanotubles Arrays for Amperometric Glucose Sensing. Adv. Funct. Mater. 2005, 15, 803–809.
Lau, K. H. A.; Tan, L. S.; Tamada, K.; Sander, M. S.; Knoll, W. Highly Sensitive Detection of Processes Occurring Inside Nanoporous Anodic Alumina Templates: A Waveguide Optical Study. J. Phys. Chem. B 2004, 108, 10812–10818.
Djokic, S. S.; Burrell, R. E. Visual Detection of Protein Adsorption onto Electrochemically Oxidized Aluminum Surfaces. Biosens. Bioelectron. 1998, 13, 271–278.
Rahmelow, K.; Hubner, W. Secondary Structure Determination of Proteins in Aqueous Solution by Infrared Spectroscopy: A Comparison of Multivariate Data Analysis Methods. Anal. Biochem. 1996, 241, 5–13.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Published online July 14, 2005
Rights and permissions
About this article
Cite this article
Wang, Y., Xia, X. & Guo, Y. Porous anodic alumina membrane as a sample support for MALDI-TOF MS analysis of salt-containing proteins. J Am Soc Mass Spectrom 16, 1488–1492 (2005). https://doi.org/10.1016/j.jasms.2005.04.014
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1016/j.jasms.2005.04.014