RJacobson_Dissertation.pdf (7.04 MB)

HILIC-MS analysis of protein glycosylation using nonporous silica

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posted on 16.01.2019 by Rachel E. Jacobson
The objective of this research is to develop and apply a HILIC UHPLC stationary phase that allows for separation of intact glycoproteins. In Chapter 1 I give an overview of the problems of current glycosylation profiling with regards to biotherapeutics, and my strategy to separate the intact glycoprotein with HILIC. Chapter 2 describes the methods used to produce the nonporous packing material and stationary phase. In Chapter 3 I describe previous work in developing a HILIC polyacrylamide stationary phase, and further improvements I have made. Chapter 4 describes development of an assay in collaboration with Genentech of therapeutic mAb glycosylation. In Chapter 5, I show HILIC-MS of digested ribonuclease B as a beginning step to analyze glycosylated biomarkers.

Funding

NIH grant 1R01GM121910-01

History

Degree Type

Doctor of Philosophy

Department

Chemistry

Campus location

West Lafayette

Advisor/Supervisor/Committee Chair

Mary Wirth

Advisor/Supervisor/Committee co-chair

Peter Kissinger

Additional Committee Member 2

Chittaranjan Das

Additional Committee Member 3

Abram Axelrod

Licence

Exports