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Document Type

Open Access Dissertation

Degree Name

Doctor of Philosophy (PhD)

Degree Program

Chemistry

Year Degree Awarded

2015

Month Degree Awarded

May

First Advisor

Igor A. Kaltashov

Subject Categories

Analytical Chemistry | Biotechnology

Abstract

Modern development of medicine requires detailed characterization by state-of-the art analytical techniques that can be used to analyze covalent structure, conformations and protein-receptor interaction to quantitatively measure biodistribution of protein therapeutics. Mass spectrometry has already become an indispensable tool facilitating all stages of protein drug development. Particularly, this work has demonstrated the tremendous potential of electrospray ionization (ESI) mass spectrometry (MS) in this arena by providing invaluable information beyond mass measurement that can be used to optimize protein drug conjugate structures during early stages of development, and to further catalyze drug design efforts. Additionally, a new sensitive and selective method that uses metal tracers and inductively coupled plasma (ICP) MS developed in our lab has been successfully applied for quantitating exogenous transferrin (Tf) and Tf-based drugs in biological tissues and fluids. Furthermore, ICP-MS based method using metal tracer in combination with size exclusion chromatography (SEC) method proved to be able to probe into protein stability post-injection and to yield useful data not accessible by other methods. For the first time a small soluble protein aggregation of injected protein drug was studied in live animals. Finally, a simple and cost-effective 18O labeling-based method has been developed for quantitating lysine modification sites of protein drug conjugates and has been successfully applied for N-succinimidyl-S-acetylthioacetate (SATA)-Lysozyme (Lz) conjugate.

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