1. Preparation and characterization of nitrilotriacetic-acid-terminated self-assembled monolayers on gold surfaces for matrix-assisted laser desorption ionization-time of flight-mass spectrometry analysis of proteins and peptides
- Author
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Andrew Vogt, Richard S. Smith, T. F. Holzman, Jianwei Shen, Robert E. Johnson, Jieyi Wang, John E. Harlan, Tanveer F. Ahmed, Sally Dorwin, and Jean M. Severin
- Subjects
Nitrilotriacetic Acid ,MALDI imaging ,Chromatography ,Protein mass spectrometry ,Surface Properties ,Chemistry ,Biophysics ,Nitrilotriacetic acid ,Proteins ,Self-assembled monolayer ,Cell Biology ,Mass spectrometry ,Biochemistry ,Recombinant Proteins ,Sample preparation in mass spectrometry ,chemistry.chemical_compound ,Matrix-assisted laser desorption/ionization ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Histidine ,Sample preparation ,Gold ,Peptides ,Molecular Biology - Abstract
On-target affinity capture, enrichment and purification of biomolecules improve detection of specific analytes from complex biological samples in matrix-assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS) analysis. In this paper, we report a simple method for preparation of a self-assembled nitrilotriacetic acid (NTA) monolayer on gold surface which can be used as a MALDI-TOF-MS sample target specifically for recombinant oligohistidine-tagged proteins/peptides and phosphorylated peptides. The NTA functional groups are immobilized to the gold surface via the linkage of 1,8-octanedithiol which forms a self-assembled monolayer on gold. Characterization by X-ray photoelectron spectroscopy and MALDI analysis of the modified surface are described. The chemically modified surface shows strong affinity toward the analytes of interest, which allows effective removal of the common interferences, e.g. salts and detergents, and therefore leads to improved signal/noise ratio and detection limit. The use of the modified surface simplifies the sample preparation for MALDI analysis of these targeted analytes.
- Published
- 2005
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