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Therapeutic Protein and Glycoprotein Production, Optimization, and Analysis Methods
Toumi, Melinda L.
Toumi, Melinda L.
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Abstract
Proteins and glycoproteins have the potential to improve health when used as therapeutic products for prevention or treatment of disease, but methods are needed for solving some remaining challenges to production, optimization, and analysis. In this dissertation, novel glycosylation engineering strategies are used to solve challenges in conventional protein production and are used to improve therapeutic protein stability by blocking asparagine deamidation, a ubiquitous cause of protein degradation. Glycan structures on therapeutic glycoproteins must be optimized to avoid negative impacts on pharmacological properties. Methods to optimize glycans are described within, using novel, extracellular glycan trimming reactions performed with glycosidases that can be implemented without harm to protein activity or stability. Finally, analysis of proteins with mass spectrometric peptide mass fingerprinting techniques can be complex, so mass defect filters used for data analysis were improved by determining the correct filter size, using experimental human protein data.
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Date
2010-05-31
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University of Kansas
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Keywords
Analytical chemistry, Pharmaceutical chemistry, Biopharmaceutics, Glycobiology, Glycoprotein, Mass spectrometry, Protein