Preparation of Heparin Surface for Quantification of Fibroblast Growth Factor-2 (FGF-2) Binding Using Surface Plasmon Resonance (SPR)

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.

Name thesis:degree_name

Master of Science

Level thesis:degree_level

masters

Discipline thesis:degree_discipline

Biomedical Engineering and Sciences

Department dc:contributor.department

Biomedical Engineering and Sciences

Grantor dc:publisher

Virginia Tech

Year dc:date.issued

2005

Author dc:creator

• Kirtland, David Rand

Chair dc:contributor.committeechair

• Forsten-Williams, Kimberly

Committee members dc:contributor.committeemember

• Davis, Richey M.
• Dessy, Raymond E.

• 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

Statement dc:rights

• In Copyright

Licence dc:rights.uri

• http://rightsstatements.org/vocab/InC/1.0/

Dc Identifier Other

etd-05262005-134908

OAI identifier oai:identifier

oai:vtechworks.lib.vt.edu:10919/33265