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

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

Bibliographic Details
Main Author: Liu, Yang
Language:English
Published: Ohio University / OhioLINK 2015
Subjects:
Biophysics
the force dependence of receptor ligand bonding
ultrasensitive force techniques
microcantilevers
the interaction of protein A and human IgG
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1420552185
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spelling ndltd-OhioLink-oai-etd.ohiolink.edu-ohiou14205521852021-08-03T06:28:51Z Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System Liu, Yang Biophysics the force dependence of receptor ligand bonding ultrasensitive force techniques microcantilevers the interaction of protein A and human IgG 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. 2015-08-25 English text Ohio University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1420552185 http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1420552185 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection NDLTD
language English
sources NDLTD
topic Biophysics
the force dependence of receptor ligand bonding
ultrasensitive force techniques
microcantilevers
the interaction of protein A and human IgG
spellingShingle Biophysics
the force dependence of receptor ligand bonding
ultrasensitive force techniques
microcantilevers
the interaction of protein A and human IgG
Liu, Yang
Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System
author Liu, Yang
author_facet Liu, Yang
author_sort Liu, Yang
title Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System
title_short Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System
title_full Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System
title_fullStr Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System
title_full_unstemmed Measurement of Force Dependence of Receptor-Ligand Bonding Using a Novel Forced Unbinding System
title_sort measurement of force dependence of receptor-ligand bonding using a novel forced unbinding system
publisher Ohio University / OhioLINK
publishDate 2015
url http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1420552185
work_keys_str_mv AT liuyang measurementofforcedependenceofreceptorligandbondingusinganovelforcedunbindingsystem
_version_ 1719437616962600960

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