Improved Methods for Structural Characterization of Therapeutic Proteins

Biosimilars, Fundamental Research, Life Sciences,
  • June 19, 2015

New Solutions and Strategies for Streamlined Glycosylation Analysis

In the areas of biotherapeutics, biomarker discovery, and glycoproteomics, it is essential to discriminate the protein and carbohydrate structures. Peptide-N-Glycosidase F (PNGase F) is a broad specificity amidase that liberates most N-linked glycans from mammalian glycoproteins. In recent years, PNGase F has become increasingly important in the biopharma industry, where defining the structure of N-glycans on therapeutic glycoproteins (e.g. antibodies) is imperative for defining their performance and quality control, and validating the structural equivalence of biosimilars. Additionally, PNGase F is increasingly being used in high throughput analyses of novel glycan-based markers of disease and development of diagnostics. The changing landscape of glycan analysis has placed new demands on the enzymes used for glycan removal with respect to their suitability for regulated environments and for high-throughput workflows.

In this webinar the presenters will introduce a series of novel products and techniques to facilitate glycoprotein analysis. First, we will present an improved reagent, Rapid PNGase F, which allows for the complete deglycosylation of therapeutic monoclonal antibodies in just minutes. Validation of complete deglycosylation with Rapid PNGase F has been demonstrated using a variety of therapeutic monoclonal antibodies. Glycan labeling and clean up workflows have been optimized, allowing for the rapid preparation of glycans for analysis by LC-MS. We will also demonstrate the utility of several novel formulations of PNGase F, including an immobilized PNGase F suitable for solid-phase applications, a PNGase F fusion that can be easily removed from workflows, and a lyophilized PNGase F with exceptional stability. In addition, techniques using alternative endoglycosidases will be reviewed, such as Endo S, for the analysis of glycosylation of monoclonal antibodies. Finally, the use of exoglycosidases will be demonstrated, alone or in combination with endoglycosidases, for the study of complex protein mixtures, and / or proteins that are heavily N– and O-glycosylated.

Speakers

Alicia Bielik, PhD, Product Manager, Glycobiology, New England Biolabs

Alicia Bielik received her PhD in Chemistry from Boston University, investigating the role of glycosaminoglycans. Specifically, her research focused on the quantification of sulfated carbohydrates in healthy and osteoarthritic cartilage using the technique of mass spectrometry. As the Group Leader of NEB’s Glycobiology & Proteomics production team, she focuses her efforts on the purification and characterization of all of the enzymes and reagents in the Glycobiology and Proteomics product portfolio. This portfolio includes over 40 products, primarily recombinant enzymes. Her efforts are focused on producing NEB’s endoglycosidase, exoglycosidase and protease products to a high quality standard, and with a level of characterization suitable for regulated environments and complex workflows.

Paula Magnelli, PhD, Scientist, Glycobiology, New England Biolabs

Paula Magnelli received her PhD from the University of Buenos Aires, Argentina, investigating the physiology of cellulose-degrading fungi. During her postdoctoral training at Boston University, she investigated the role and structure of glycoconjugates of the yeast cell wall and of pathogenic protozoans. While characterizing unknown glycans she became aware of the limitations imposed by an inadequate toolkit to study carbohydrate structure. As a Scientist within the Glycobiology Team at NEB, Dr. Magnelli first worked on the discovery and characterization of novel enzymes, to expand the repertoire of available tools for Glycobiology research. In her current role, she integrates chemical and enzymatic methods to optimize sample preparation workflows for glycomics and glycoproteomics. Her research incorporates concepts of quality by design, creating innovative techniques that are robust, user friendly, and can be integrated in current workflows.

Who Should Attend?

Biopharma professionals working within:

  • Biosimilars
  • Therapeutic monoclonal antibodies
  • Product development & manufacturing
  • Quality control

The presentation will also be beneficial for Contract Research Organizations performing glycan analysis.

Xtalks Partners

New England Biolabs

Established in the mid 1970’s, New England Biolabs, Inc. (NEB) is the industry leader in the discovery and production of enzymes for molecular biology applications and now offers the largest selection of recombinant and native enzymes for genomic research. NEB continues to expand its product offerings into areas related to PCR, gene expression, sample preparation for next generation sequencing, synthetic biology, glycobiology, epigenetics and RNA analysis. Additionally, NEB is focused on strengthening alliances that enable new technologies to reach key market sectors, including molecular diagnostics development. New England Biolabs is a privately held company, headquartered in Ipswich, MA, and has extensive worldwide distribution through a network of exclusive distributors, agents and seven subsidiaries located in Canada, China, France, Germany, Japan, Singapore and the UK. For more information about New England Biolabs visit www.neb.com.

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