Development Of Matrix Assisted Laser Desorption Ionization And Liquid Chromatography Electrospray Ionization Based Mass Spectrometric Techniques For Characterizing And Quantifying Endogenous And Therapeutic Biomolecules
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Development of Matrix-assisted Laser Desorption/ionization and Liquid Chromatography - Electrospray Ionization Based Mass Spectrometric Techniques for Characterizing and Quantifying Endogenous and Therapeutic Biomolecules
Author | : Bingming Chen |
Publisher | : |
Total Pages | : 463 |
Release | : 2017 |
Genre | : |
ISBN | : |
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Endogenous biomolecules, such as neuropeptide, protein and carbohydrates, play important roles in biological and physiological processes. Over the last two decades, therapeutic biomolecules, including therapeutic peptides, proteins and monoclonal antibodies, have become large and fast-growing segments of the pharmaceutical industry. However, analyzing biomolecules is challenging due to their large size, complex structure and various modifications. In situ analysis of biomolecules is even more challenging due to the large dynamic range and complex microenvironments on tissue surface. In this dissertation, matrix-assisted laser desorption/ionization (MALDI) and liquid chromatography (LC) -- electrospray ionization (ESI) based mass spectrometric techniques were developed to characterize and quantify endogenous and therapeutic biomolecules. Specifically, this work developed novel ionization techniques and mass spectrometry imaging workflows to study the sequence and in situ distribution of biomolecules on high resolution accurate mass instrument platforms. Moreover, absolute or relative quantifications of biomolecules, such as therapeutic peptide or N-glycans, were measured by cutting-edge quantification workflows. In summary, this work not only improves current analytical methodology for characterizing biomolecules, but also demonstrates its potential applications in pharmaceutical industry and clinical settings.
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