Description

In this course you will learn the fundamentals of carbohydrate chemistry and their analysis. We review the types of glycoconjugates in mammalian cells and their recognized biological functions. We describe glycan biosynthesis as a way to understand carbohydrate heterogeneity and discuss the diversity of N- and O-glycan structures in humans in comparison to those expressed by other organisms. We review the structures that are common to different types of glycans and the physicochemical methods used for their characterization. We discuss the strategies for method validation and setting specifications. We review the evaluation of critical structural features at early stages to aid in the selection of adequate cell lines and process parameters resulting in the desired product quality profile. The objective is to provide participants with key technical information along with perspectives to enable them to evaluate the carbohydrates present in their own products and confidently select analytical strategies to support the various stages of product development.

Agenda

• The Basis for Structural Complexity in Carbohydrates
  • stereochemistry of simple sugars
  • monosaccharides: types, relationship, properties
  • chemical reactions of monosaccharides
  • monosaccharides as building blocks: the glycosidic linkage
  • oligosaccharides and polysaccharides: The glycosidic linkage impact on molecular shape and interactions
• Carbohydrates in Mammalian Cells
  • types of glycoconjugates
  • structural and modulatory function of glycans
  • the heparin-antithrombin binding requirements
• Understanding Carbohydrate Heterogeneity
  • overview of carbohydrate biosynthesis
  • biosynthesis of N- and O-glycans
  • glycans in evolution: glycans expressed in bacteria, yeast, insects and plants
  • structures common to different types of glycans
  • monitoring sialic acids as sensitive stability indicators
• Analysis of Carbohydrate Structures
  • defining the analytical needs for each stage of development
  • identifying the critical reporter features of the expected glycan structures
  • selecting the analytical tool box: glycosylation analysis platform
  • analysis of carbohydrates in glycoproteins: isoforms; site-specific glycosylation; oligosaccharide profiling; monosaccharide composition; sialic acids
  • overview of glycosaminoglycan (GAG) analysis
• Complete Characterization of Carbohydrate Structures
  • linkage analysis: permethylation
  • sequence analysis: NMR vs. mass spectrometry; exoglycosidases

Who Should Attend?

This course is designed for Scientists and Managers in the life science industry whose job functions involve analytical methods development in support of upstream and downstream in-process monitoring, formulation, quality control, stability studies and research and development of any type of carbohydrate-containing biopharmaceutical.

Instructor

Adriana E. Manzi, Ph.D., President and Principal Consultant, Manzi & Associates
Adriana E. Manzi, Ph.D. has lectured on carbohydrate chemistry since 1980, first in Argentina, then as Assistant Professor at the University of California San Diego. She has published 28 original research articles and 20 book chapters in the field of carbohydrate analysis. Dr. Manzi's work has encompassed the technical, quality, and regulatory aspects of CMC studies necessary to achieve biotechnology product approval. She led biomolecule structural analysis teams with focus on physicochemical methods supporting all stages of product development. Dr. Manzi has been Associate Director of Analytical Development at Cytel (carbohydrate therapeutics), Director of Analytical Development/QC at Nextran (xenotransplantation) and Senior Director of Research at Baxter Healthcare. In 2006, she founded Manzi & Associates and is now consulting for biopharmaceuticals R&D.