Analysis and Applications of Fluorescence Fluctuation Spectroscopy
Chen, Yan
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https://hdl.handle.net/2142/85488
Description
Title
Analysis and Applications of Fluorescence Fluctuation Spectroscopy
Author(s)
Chen, Yan
Issue Date
1999
Doctoral Committee Chair(s)
Gratton, Enrico
Department of Study
Biophysics and Computational Biology
Discipline
Biophysics and Computational Biology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Statistics
Language
eng
Abstract
Binary dye mixtures were used to demonstrate the influence of the molecular brightness on the fluctuation amplitude G(0). This dependence allows to resolve multiple species based on the difference in their molecular brightness, which we applied to a titration study of an antibody with its ligand. The global fit of the average photon counts 〈k〉 and the fluctuation amplitude G(0) as a function of the protein concentration determined the number of binding sites and the dissociation coefficient KD. The analysis further demonstrated heterogeneity of the antibody complex on a time scale not easily accessible by other techniques.
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