Development of novel analytical tools for complex carbohydrates

Complex carbohydrates such as the heparin/heparan sulfate glycosaminoglycans (HSGAGs) and the asparagine linked glycans (N-glycans) have emerged as crucial components of many human biological systems. Both classes of molecules have diverse roles in normal and pathological processes including development, cell growth, wound healing, angiogenesis, viral entry, and cancer. Taking advantage of the ability of carbohydrates to affect biological outcomes, several companies have already begun developing sugar-based structures as therapeutics for malaria and breast cancer, among others. However, the structural complexity that allows HSGAGs and N-glycans to exhibit such a wide array of activities also makes them more difficult to study than their protein and nucleotide counterparts. There are currently no effective methods for rapid synthesis of specific HSGAG or N-glycan structures, and until recently, most common analytical techniques were not useful in this field due to sensitivity constraints. Now, with advances in separation techniques and mass spectrometry, the fundamental roles of biological carbohydrates can be elucidated more efficiently. However, there is still a need for improved methodologies for studying these classes of molecules. This thesis describes the development of a novel technique for the direct isolation and sequencing of protein-binding HSGAG oligosaccharides. As a proof-of-concept, we used heparin and HSGAG-binding proteins of known specificity (fibroblast growth factor and antithrombin III), then utilized the technique to identify an oligosaccharide binder for a protein with unknown HSGAG binding specificity (endostatin). Furthermore, this technique can be readily applied to carbohydrate material from the cell surface. We have

Department dc:contributor.department

Massachusetts Institute of Technology. Biological Engineering Division.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2004

Author dc:creator

• Keiser, Nishla, 1978-

Advisor dc:contributor.advisor

• Ram Sasisekharan.

• Biological Engineering Division.

Statement dc:rights

• MIT theses are protected by copyright. They may be viewed, downloaded, or printed from this source but further reproduction or distribution in any format is prohibited without written permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

en_US

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/28525

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/28525