Title
Synthesis of an Affinity Ligand for the Separation of Glycosylated and Non-Glycosylated Proteins
Primary Submission Contact
Laura Rowe
Faculty Sponsor
Laura Rowe
Faculty Sponsor Email Address
laura.rowe@valpo.edu
College
Arts and Sciences
Department/Program
Chemistry
Document Type
Poster Presentation
Date
Fall 10-28-2016
Abstract
When studying glycosylated proteins, a method of separating glycosylated proteins from other proteins is crucial for purifying the sample. Affinity chromatography is currently performed most commonly for glycoprotein purification, through the use of boronic acid based, or lectin derived, ligands. However, more specific and economical ligands would be preferred. One such ligand described here is an affinity ligand that binds to glycosylated proteins and the monosaccharide α-D-mannopyranoside, synthesized through an Ugi-multicomponent reaction. The ligand is composed of tryptamine, 1-benzyl-1H-indole-3-carboxylic acid, ispropyl isocyanide, and an aldehyde-functionalized Sepharose bead. The ligand’s structure was selected via performance in a previously produced combinatorial library. For this work, after aldehyde functionalizing Sepharose beads and completing the Ugi reaction, the binding ability of the ligand is tested with gravity packed chromatography columns using glucose oxidase as the protein of interest. Then a Bradford assay is conducted in order to determine the amount of glycosylated protein (glucose oxidase) that bound to the affinity ligand during a positive and negative screening process. The ligand was synthesized once and tested with inconclusive results, and therefore resynthesis has been completed.
Recommended Citation
Gunter, Amy J. and Rowe, Laura, "Synthesis of an Affinity Ligand for the Separation of Glycosylated and Non-Glycosylated Proteins" (2016). Fall Interdisciplinary Research Symposium. 10.
https://scholar.valpo.edu/fires/10
Biographical Information about Author(s)
Amy Gunter is a junior biochemistry major at Valparaiso University. She intends to attend graduate school and pursue a career in the realm of biochemistry and organic chemistry research.