Date of Award

Spring 2022

Degree Type

Thesis

Department

Chemistry and Biochemistry

Director of Thesis

Dr. Qian Wang

Second Reader

Jingyu Fan

Abstract

Streptavidin, a bacteria-derived protein, and the small molecule biotin display extremely high-affinity binding. Researchers have taken advantage of this uniquely strong interaction in a myriad of detection, labeling, and purification techniques, one of which is affinity chromatography. Streptavidin can be immobilized onto a solid support, and this streptavidin-conjugated material can be used to capture biotin-tagged molecules with exceedingly high strength and specificity. In this project, a protocol was developed for synthesizing an agarose-based streptavidin resin for use as an affinity chromatography medium. Additionally, two fluorescence-based assays were developed for quantifying the amount of streptavidin conjugated to each resin and for quantifying the ability of each resin to capture biotin-tagged biomolecules. Several streptavidin resins were synthesized with varied reaction parameters in an attempt to maximize the resin’s ability to capture biotin-tagged molecules. The result of this project is a streptavidin resin with optimized biotin-binding capabilities that has the potential to be implemented as a versatile tool for selectively capturing biomolecules of interest.

First Page

1

Last Page

27

Available for download on Wednesday, May 01, 2024

Included in

Biotechnology Commons

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