Title page for ETD etd-05262005-134908


Type of Document Master's Thesis
Author Kirtland, David Rand
Author's Email Address drk@vt.edu
URN etd-05262005-134908
Title Preparation of Heparin Surface for Quantification of Fibroblast Growth Factor-2 (FGF-2) Binding Using Surface Plasmon Resonance (SPR).
Degree Master of Science
Department Biomedical Engineering and Sciences
Advisory Committee
Advisor Name Title
Forsten-Williams, Kimberly Committee Chair
Davis, Richey M. Committee Member
Dessy, Raymond E. Committee Member
Keywords
  • Fibroblast Growth Factor-2 (FGF-2) basic Fibroblas
  • Mixed Self-Assembled Monolayer
  • Mixed Self Assembling Monolayer (mSAM)
  • Surface Plasmon Resonance (SPR)
  • Growth Factor
  • Biosensor
  • Heparin
Date of Defense 2005-05-04
Availability unrestricted
Abstract
A mixed self assembling monolayer (mSAM) chip with attached heparin was developed to analyze heparin-protein interactions using a Reichert Inc, SR7000, surface plasmon resonance (SPR) instrument. The heparin was attached via streptavidin-biotin linkage where the streptavidin was covalently coupled to the mSAM and biotinylated heparin bound to it. These chips were then used to quantify the interactions of fibroblast growth factor-2 (FGF-2) with the surface bound heparin. Kinetic rate constants of association and disassociation were calculated. The association data of FGF-2 with heparin was fit to a single compartment, well-mixed model as the data did not exhibit mass transfer limitations. The results suggested that rebinding was prevalent and observed disassociation rates differed significantly in the presence of competing soluble heparin during disassociation. Our results indicate that the Reichert instrument and mSAM chips can be used to analyze heparin-protein interactions but that a careful protocol, outlined in this thesis, should be followed to obtain optimal data.
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