Citation
Baio, J. E.; Weidner, T.; Samuel, N. T.; McCrea, Keith; Baugh, Loren; Stayton, Patrick S.; & Castner, David G. (2010).
Multi-technique Characterization of Adsorbed Peptide and Protein Orientation: LK310 and Protein G B1.
Journal of vacuum science & technology. B, Microelectronics and nanometer structures : processing, measurement, and phenomena : an official journal of the American Vacuum Society, 28(4).
Abstract
The ability to orient biologically active proteins on surfaces is a major challenge in the design, construction, and successful deployment of many medical technologies. As methods to orient biomolecules are developed, it is also essential to develop techniques that can an accurately determine the orientation and structure of these materials. In this study, two model protein and peptide systems are presented to highlight the strengths of three surface analysis techniques for characterizing protein films: time-of-flight secondary ion mass spectrometry (ToF-SIMS), sum-frequency generation (SFG) vibrational spectroscopy, and near-edge x-ray absorption fine structure (NEXAFS) spectroscopy. First, the orientation of Protein G B1, a rigid 6 kDa domain covalently attached to a maleimide-functionalized self-assembled monolayer, was examined using ToF-SIMS. Although the thickness of the Protein G layer was similar to the ToF-SIMS sampling depth, orientation of Protein G was successfully determined by analyzing the C2H5S+ intensity, a secondary ion derived from a methionine residue located at one end of the protein. Next, the secondary structure of a 13-mer leucine-lysine peptide (LK310) adsorbed onto hydrophilic quartz and hydrophobic fluorocarbon surfaces was examined. SFG spectra indicated that the peptide’s lysine side chains were ordered on the quartz surface, while the peptide’s leucine side chains were ordered on the fluorocarbon surface. NEXAFS results provided complementary information about the structure of the LK310 film and the orientations of amide bonds within the LK310 peptide.
Reference Type
Journal Article
Secondary Title
Journal of vacuum science & technology. B, Microelectronics and nanometer structures : processing, measurement, and phenomena : an official journal of the American Vacuum Society
Author(s)
Baio, J. E.Weidner, T.Samuel, N. T.McCrea, KeithBaugh, LorenStayton, Patrick S.Castner, David G.
Year Published
2010
Date Published
1277942400
Volume Number
28
Issue Number
4
ISSN/ISBN
1071-1023
DOI
Multi-technique Characterization of Adsorbed Peptide and Protein Orientation10.1116/1.3456176
