Date of Award
5-2024
Document Type
Thesis
Degree Name
Master of Science (MS)
Department
Chemistry
Committee Chair/Advisor
Dr. R. Kenneth Marcus
Committee Member
Dr. Terri F. Bruce
Committee Member
Dr. Carlos D. Garcia
Committee Member
Dr. Daniel C. Whitehead
Abstract
In the past two decades, Marcus and colleagues have led the advancement of capillary-channeled polymer (C-CP) fiber stationary phases for biomacromolecule separations, including proteins and nanovesicles. These stationary phases offer notable advantages, including low cost ($5/column), straightforward column fabrication, and the capacity for on-column surface modification, enabling high chemical selectivity. This study examines a C-CP fiber modification strategy for capturing target proteins and develops a Tween-20-assisted C-CP-based hydrophobic interaction chromatography (HIC) method for the isolation and quantification of exosomes from human urine.
The quest for higher-throughput and cost-effective protein isolation techniques remains a key focus in a variety of end uses. The streptavidin-biotin interaction, known for its exceptional specificity and stability, offers a promising avenue for protein purification. In this approach, immobilization of streptavidin (SAV) onto a solid substrate is crucial for the effective capture of biotinylated proteins. This study explores the potential of polypropylene (PP) C-CP fibers as support phases for SAV immobilization via direct adsorption. The PP C-CP fiber columns modified with SAV efficiently capture biotinylated proteins, indicating the promise of this approach for fast and cost-effective target protein capture.
Exosomes, membrane-secreted vesicles ranging from 30 to 200 nm, play crucial roles in intercellular communication and hold potential as therapeutic delivery vehicles. Efficient isolation and quantification methods are essential for studying their functions and therapeutic applications. Marcus and coworkers have developed Polyester (PET), capillary-channeled polymer fiber columns, and spin-down tips as high-throughput and cost-effective approaches for exosome isolation. However, selecting the optimal elution solvent remains a challenge, particularly for preserving the structural integrity of the isolated vesicles. This study evaluates Tween-20, a non-ionic detergent, as an elution solvent for exosome recovery from human urine in advancing the versatility of C-CP fiber column-based HIC isolation methods.
Recommended Citation
Islam, Md Khalid Bin, "Biotinylated Protein Capture and Tween-20 Assisted Exosome Isolations via Capillary Chanelled Polymer Fiber" (2024). All Theses. 4249.
https://tigerprints.clemson.edu/all_theses/4249
Author ORCID Identifier
https://orcid.org/0000-0001-9029-6092