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Enhanced TOF-SIMS imaging of a micropatterned protein by stable isotope protein labeling.

Publication ,  Journal Article
Belu, AM; Yang, Z; Aslami, R; Chilkoti, A
Published in: Analytical chemistry
January 2001

Patterning of biomolecules on surfaces is an increasingly important technological goal. Because the fabrication of biomolecule arrays often involves stepwise, spatially resolved derivatization of surfaces, spectroscopic imaging of these arrays is important in their fabrication and optimization. Although imaging time-of-flight secondary ion mass spectrometry (TOF-SIMS) is a powerful method for spatially resolved surface analysis, TOF-SIMS images of micropatterned proteins on organic substrates can be difficult to acquire, because of the lack of high intensity, protein-specific molecular ions that are essential for imaging under static conditions. In contrast, low-mass ions are of suitable intensity for imaging, but can originate from different chemical species on the surface. A potential solution to this problem is to utilize stable isotope labeled proteins, an approach that has heretofore not been explored in TOF-SIMS imaging of micropatterned proteins and peptides. To investigate the feasibility of stable isotope enhanced TOF-SIMS imaging of proteins, we synthesized 15N-labeled streptavidin by labeling of the protein during expression from a recombinant gene. The spatial distribution of streptavidin bound to biotin micropatterns, fabricated on a polymer and on a self-assembled monolayer on gold, was imaged by TOF-SIMS. Imaging of high-intensity, low-m/z secondary ions (e.g., C15N-) unique to streptavidin enabled unambiguous spatial mapping of the micropatterned protein with a lateral resolution of a few micrometers. TOF-SIMS imaging of micropatterned 15N-labeled streptavidin also illustrated the exquisite sensitivity of TOF-SIMS to low fractional coverage of protein (5 A effective thickness) in the background regions of the protein micropattern.

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Published In

Analytical chemistry

DOI

EISSN

1520-6882

ISSN

0003-2700

Publication Date

January 2001

Volume

73

Issue

2

Start / End Page

143 / 150

Related Subject Headings

  • Streptavidin
  • Recombinant Proteins
  • Mass Spectrometry
  • Image Processing, Computer-Assisted
  • Analytical Chemistry
  • 4004 Chemical engineering
  • 3401 Analytical chemistry
  • 3205 Medical biochemistry and metabolomics
  • 0399 Other Chemical Sciences
  • 0301 Analytical Chemistry
 

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Belu, A. M., Yang, Z., Aslami, R., & Chilkoti, A. (2001). Enhanced TOF-SIMS imaging of a micropatterned protein by stable isotope protein labeling. Analytical Chemistry, 73(2), 143–150. https://doi.org/10.1021/ac000771l
Belu, A. M., Z. Yang, R. Aslami, and A. Chilkoti. “Enhanced TOF-SIMS imaging of a micropatterned protein by stable isotope protein labeling.Analytical Chemistry 73, no. 2 (January 2001): 143–50. https://doi.org/10.1021/ac000771l.
Belu AM, Yang Z, Aslami R, Chilkoti A. Enhanced TOF-SIMS imaging of a micropatterned protein by stable isotope protein labeling. Analytical chemistry. 2001 Jan;73(2):143–50.
Belu, A. M., et al. “Enhanced TOF-SIMS imaging of a micropatterned protein by stable isotope protein labeling.Analytical Chemistry, vol. 73, no. 2, Jan. 2001, pp. 143–50. Epmc, doi:10.1021/ac000771l.
Belu AM, Yang Z, Aslami R, Chilkoti A. Enhanced TOF-SIMS imaging of a micropatterned protein by stable isotope protein labeling. Analytical chemistry. 2001 Jan;73(2):143–150.
Journal cover image

Published In

Analytical chemistry

DOI

EISSN

1520-6882

ISSN

0003-2700

Publication Date

January 2001

Volume

73

Issue

2

Start / End Page

143 / 150

Related Subject Headings

  • Streptavidin
  • Recombinant Proteins
  • Mass Spectrometry
  • Image Processing, Computer-Assisted
  • Analytical Chemistry
  • 4004 Chemical engineering
  • 3401 Analytical chemistry
  • 3205 Medical biochemistry and metabolomics
  • 0399 Other Chemical Sciences
  • 0301 Analytical Chemistry