Effect of fluorescently labeling protein probes on kinetics of protein-ligand reactions

Y. S. Sun, J. P. Landry, Y. Y. Fei, X. D. Zhu, J. T. Luo, X. B. Wang, Kit Lam

Research output: Contribution to journalArticle

75 Citations (Scopus)

Abstract

Westudied the effect of fluorescently labeling proteins on protein-ligand reactions. Unlabeled ligands (streptaVidinbinding peptides and rabbit immunoglobulinG(IgG) as antigen targets) are immobilized on epoxy-functionalized glass slides. Unlabeled and Cy3-labeled protein probes from the same batch (streptavidin and goat antibodies) subsequently react with the surface-immobilized targets. By monitoring in situ the surface mass density change using an obliqueincidence reflectivity difference scanning microscope (a label-free detector), we measured k on and k off for streptavidin-peptide reactions and antibody-antigen reaction. We found that (1) equilibrium dissociation constants, defined as KD ) k off/k on, for streptavidin-peptide reactions increases by a factor of 3-4 when the solution-phase streptavidin is labeled with Cy3 dye and (2) K D for reactions of solution-phase goat anti-rabbit antibodies with rabbit IgG targets also change significantly when the goat antibodies are labeled with Cy3 dye.

Original languageEnglish (US)
Pages (from-to)13399-13405
Number of pages7
JournalLangmuir
Volume24
Issue number23
DOIs
StatePublished - Dec 2 2008

Fingerprint

Streptavidin
Antibodies
Labeling
Peptides
marking
goats
antibodies
Ligands
proteins
Proteins
rabbits
ligands
Kinetics
probes
kinetics
peptides
Dyes
Antigen-antibody reactions
antigens
Antigens

ASJC Scopus subject areas

  • Electrochemistry
  • Condensed Matter Physics
  • Surfaces and Interfaces
  • Materials Science(all)
  • Spectroscopy

Cite this

Sun, Y. S., Landry, J. P., Fei, Y. Y., Zhu, X. D., Luo, J. T., Wang, X. B., & Lam, K. (2008). Effect of fluorescently labeling protein probes on kinetics of protein-ligand reactions. Langmuir, 24(23), 13399-13405. https://doi.org/10.1021/la802097z

Effect of fluorescently labeling protein probes on kinetics of protein-ligand reactions. / Sun, Y. S.; Landry, J. P.; Fei, Y. Y.; Zhu, X. D.; Luo, J. T.; Wang, X. B.; Lam, Kit.

In: Langmuir, Vol. 24, No. 23, 02.12.2008, p. 13399-13405.

Research output: Contribution to journalArticle

Sun, YS, Landry, JP, Fei, YY, Zhu, XD, Luo, JT, Wang, XB & Lam, K 2008, 'Effect of fluorescently labeling protein probes on kinetics of protein-ligand reactions', Langmuir, vol. 24, no. 23, pp. 13399-13405. https://doi.org/10.1021/la802097z
Sun YS, Landry JP, Fei YY, Zhu XD, Luo JT, Wang XB et al. Effect of fluorescently labeling protein probes on kinetics of protein-ligand reactions. Langmuir. 2008 Dec 2;24(23):13399-13405. https://doi.org/10.1021/la802097z
Sun, Y. S. ; Landry, J. P. ; Fei, Y. Y. ; Zhu, X. D. ; Luo, J. T. ; Wang, X. B. ; Lam, Kit. / Effect of fluorescently labeling protein probes on kinetics of protein-ligand reactions. In: Langmuir. 2008 ; Vol. 24, No. 23. pp. 13399-13405.
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