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Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System

Liu, Yang
http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1420552185

Abstract Details

2015, Doctor of Philosophy (PhD), Ohio University, Physics and Astronomy (Arts and Sciences).
The force dependence of receptor–ligand bonding has been extensively studied in recent years. Many ultrasensitive force techniques have been used to measure receptor-ligand bonding interaction in biophysics. Microcantilevers have easily tunable and variable spring constants over other techniques. This work presents in detail the measurement methods of glass fiber microcantilever spring constants. Microcantilevers made from E-glass fibers were mounted in viewing chambers and imaged under a microscope. Multiple sets of images for each microcantilever were captured and analyzed using custom LabVIEW IMAQ image processing program with the centroid tracking images processing. The centroid data were processed using a 20 point moving average filter which was the optimum number of average points in a moving average filter. The results of microcantilever spring constants from thermal fluctuation method measurement were compared with elasticity theory calculation. The measured spring constants of 3 - 7 mm-long fibers were within the relative error expected for the spring constant due to the fiber length and diameter uncertainties. A novel forced unbinding system was developed for the receptor-ligand binding experiments. The system applied hydrodynamic flow from one micropipette to a glass fiber microcantilever tip which was brought into contact with a bead held in another micropipette with suction pressure. The displacement of the fiber tip increased linearly as the water pressure difference increased which is the magnitude of applied pressure used to generate flow from a micropipette. The interaction of protein A and human IgG was measured using the novel forced unbinding system. Heat treatment of the Bovine Serum Albumin is necessary to reduce the frequency of nonspecific adhesion. The percentage of adhesive events decreased from 2.69 % to 0.57 % after the BSA was denatured. The protein A and human IgG binding interaction was investigated for specific adhesion experiments. The lifetime as a function of force was directly measured by the method that allowed near-instantaneous application of forces. The response of protein A-human IgG bonds to force agreed best with catch-slip transition mechanics but the Bell model could not be excluded without further experiments.
David Tees (Advisor)
170 p.
Biophysics
the force dependence of receptor ligand bonding; ultrasensitive force techniques; microcantilevers; the interaction of protein A and human IgG

Recommended Citations

Citations

  • Liu, Y. (2015). Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System [Doctoral dissertation, Ohio University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1420552185

    APA Style (7th edition)

  • Liu, Yang. Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System . 2015. Ohio University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1420552185.

    MLA Style (8th edition)

  • Liu, Yang. "Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System ." Doctoral dissertation, Ohio University, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1420552185

    Chicago Manual of Style (17th edition)

ohiou1420552185
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© 2015, all rights reserved.
This open access ETD is published by Ohio University and OhioLINK.