Date of Award

2010

Degree Type

Dissertation

Department

Chemistry

First Advisor

Sun, Xue-Long

Subject Headings

Glycomics, Proteomics, Ligands (Biochemistry)

Abstract

Cell surface carbohydrates existing as parts of glycoproteins, glycolipids, and other conjugates present the first information about cell to the outside world and are intimately involved in various biological events such as intercellular communication, and molecular and cellular targeting. However, mechanisms of most processes at the molecular level are still unclear. Therefore, it is very important to develop carbohydrate-specific binding molecules for rapid, efficient, sensitive purification and accurate analysis of complex carbohydrate structures as well as their functions. Furthermore, carbohydrate-specific binding molecules can be expected to be used in medical diagnostic applications for carbohydrate biomarkers. In this thesis study, oriented and multivalent carbohydrate-binding macromolecules were designed and developed based on a chain-end functionalized boronic acid-containing polymer (boropolymer) for glyco-capturing, glycomics and glycoproteomics applications. Namely, a biotin chain end and O-cyanate chain-end functionalized boropolymers were synthesized via aryalamine initiated cyanoxyl-mediated free radical polymerization in a one-pot fashion. The resultant boropolymers were characterized by 1H-NMR and 13C NMR spectroscopy. In our first study we demonstrated the efficient glyco-capturing followed by direct MALDI mass spectrometry identification of the captured carbohydrate by using magnetic beads functionalized with the biotin boropolymer via streptavidin/biotin interaction. In our second study we demonstrated oriented and covalent immobilization of O-Cyanate chain-end functionalized boropolymer on to amine-modified solid surfaces and its specific glyco-capturing capacity by QCM and AFM techniques. We further studied the multivalent interactions of the immobilized O-Cyanate chain end functionalized boropolymer with five different carbohydrate conjugated AuNPs. Our studies showed that different carbohydrates have different binding constants. Furthermore, the multivalent binding between carbohydrates

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