YorkSpace has migrated to a new version of its software. Access our Help Resources to learn how to use the refreshed site. Contact diginit@yorku.ca if you have any questions about the migration.
 

Improving the Accuracy, Versatility, and Partitioning Efficiency of Nonequilibrium Capillary Electrophoresis of Equilibrium Mixtures

Simple item page
dc.contributor.advisorKrylov, Sergey N.
dc.creatorKanoatov, Mirzo
dc.date.accessioned2017-07-27T12:38:18Z
dc.date.available2017-07-27T12:38:18Z
dc.date.copyright2016-11-14
dc.date.issued2017-07-27
dc.date.updated2017-07-27T12:38:17Z
dc.degree.disciplineBiology
dc.degree.levelDoctoral
dc.degree.namePhD - Doctor of Philosophy
dc.description.abstractStudying the affinity and the kinetics of biomolecular interactions is an essential task in biology and pharmacology. Nonequilibrium capillary electrophoresis of equilibrium mixtures (NECEEM) is a promising technique that allows the measurement of the equilibrium dissociation constant (Kd), and the rate constant of dissociation (koff), and can also be used as the partitioning method in selection of aptamers. There are three critical issues that have hindered the wide adoption of NECEEM by the analytical community: (1) poor resilience of the method against systematic errors, (2) limited versatility in terms of emulating physiological conditions, and (3) restricted partitioning efficiency in DNA aptamer selection. The goals of my research project were (i) to develop strategies for minimizing the systematic errors in NECEEM; (ii) to make NECEEM compatible with the use of physiological buffers; and (iii) to eliminate the restricted partitioning efficiency of NECEEM in aptamer selection. To improve the accuracy of NECEEM, I have developed an approach for simultaneous determination of both the Kd and the concentration of one of the interacting molecules, which eliminates errors caused by the presence of improperly folded molecules in samples. Further, I have developed an algorithmic approach for optimization of NECEEM, which takes the interrelation between its experimental parameters into account and minimizes the systematic error in an objective manner. To make NECEEM compatible with physiological buffers, I have uncovered the cause behind the poor detectability of DNA in phosphate buffered saline, and have created a pressure-assisted modification of NECEEM that overcomes this problem. Lastly, I have determined that the irregular electrophoretic migration of DNA in strong electric fields is caused by the ability of DNA to form unusually stable complexes with its counterions. Based on these findings, I have developed two independent approaches that increase the efficiency of NECEEM in selection of DNA aptamers.
dc.identifier.urihttp://hdl.handle.net/10315/33444
dc.language.isoen
dc.rightsAuthor owns copyright, except where explicitly noted. Please contact the author directly with licensing requests.
dc.subjectBiochemistry
dc.subject.keywordsBiomolecular interactions
dc.subject.keywordsKinetics
dc.subject.keywordsAffinity
dc.subject.keywordsKinetic capillary electrophoresis
dc.subject.keywordsAptamers
dc.titleImproving the Accuracy, Versatility, and Partitioning Efficiency of Nonequilibrium Capillary Electrophoresis of Equilibrium Mixtures
dc.typeElectronic Thesis or Dissertation

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Kanoatov_Mirzo_2016_Phd.pdf
Size:
3.83 MB
Format:
Adobe Portable Document Format
Download
License bundle
Now showing 1 - 2 of 2
No Thumbnail Available
Name:
license.txt
Size:
1.83 KB
Format:
Plain Text
Description:
Download
No Thumbnail Available
Name:
YorkU_ETDlicense.txt
Size:
3.38 KB
Format:
Plain Text
Description:
Download

Collections

Biology

All items in the YorkSpace institutional repository are protected by copyright, with all rights reserved except where explicitly noted.