RJacobson_Dissertation.pdf (7.04 MB)
Download fileHILIC-MS analysis of protein glycosylation using nonporous silica
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 WirthAdvisor/Supervisor/Committee co-chair
Peter KissingerAdditional Committee Member 2
Chittaranjan DasAdditional Committee Member 3
Abram AxelrodUsage metrics
Categories
Keywords
hplclcmsuplcuhplcliquid chromatographyHILIChydrophilic interaction liquid chromatographybiologicsmAbsantibodiesproteinglycanglycosylationglycoformMSmass spectrometryAGETATRPsurface-initiatedSeparation ScienceProteins and PeptidesColloid and Surface ChemistryCharacterisation of Biological Macromolecules