CONFERENCE SERIES: Biological Therapeutic Products
Recorded at: PEGS: The Essential Protein Engineering Summit
Digital Course: Biological Mass Spectrometric Applications For Drug Discovery And Product Development
About this Product:
This digital course is geared to those individuals supporting biopharmaceutical drug characterization analyses using mass spectrometry in either discovery or product development. Aspects that will be covered in the course include:
• A novel method for assessing disulfide bond networks
• Methods for performing glycosylation analyses
• Methods for conducting PTM analyses
• Methods for performing stress assessments on drug candidates
About this Product:
Over 100 Minutes
Site License: $1380
Agenda At A Glance:
Quantitation of Released Glycoprotein Glycans
Ron Orlando, Ph.D., Professor, Biochemistry and Molecular Biology, Complex Carbohydrate Research Center, University of Georgia
One of the analytical challenges facing scientists in the characterization of glycoproteins involves the ability to quantify changes in the attached glycans. Multiple strategies are currently utilized to accomplish this task, including; both label-free and isotopic labeling strategies. The focus of this talk will be to describe each of these approaches, and provide insight into their strengths and weaknesses, thus allowing investigators to make an educated decision on the strategy that is best suited for their particular application.
Biography: Dr. Orlando has over 25 years of experience with mass spectrometry, 21 of these years focused on the identification/characterization of proteins and their post-translational modifications (particularly glycosylation), and has published over 100 articles in scientific journals along with numerous book chapters. Additionally, Dr. Orlando is the founder and current Chair of the Associations of Biomolecular Resource Facilities’ Glycoprotein Research Group.
Application of MALDI TOF/TOF Mass Spectrometry and Collision-Induced Dissociation for the Rapid Identification of Disulfide-Bonded Peptides
Dariusz Janecki, Ph.D., Research Scientist, Biologics Mass Spectrometry & Allied Technologies, Centocor R&D, Inc.
One of the challenges in characterizing proteins is post-translational modifications. A MALDI TOF-TOF MS method has been developed for the quick and efficient identification of disulfide-bonded peptides directly from native digestions without pre-separation steps. Due to the fragility of disulfide bonds, few peptide backbone ions are produced; however, distinctive “disulfide bond triplet” ion signatures are produced that can be used to identify the presence and composition of disulfide-bonded peptides. The method provides significant help in mapping disulfide-bond networks.
Biography: Dr. Janecki entered the biopharmaceutical industry in 2004 as a research scientist at a startup biotech company and in 2008, joined Centocor’s Discovery Research department mass spectrometry group which focuses on the characterization of molecules from inception through pre-clinical. The principal role of the group is structural characterization of biopharmaceuticals, and the antigens and receptors used during drug development. Dr. Janecki obtained his Ph.D. degree in Analytical Chemistry from Jagiellonian University (Cracow, Poland) in 2001, and subsequently accepted a post-doctoral research fellowship at Indiana University in Bloomington, Indiana (group of Prof. James P. Reilly), during which he focused on proteomics and applied bioanalytical mass spectrometry. His publications include protein characterizations, as well as methods for qualitative and quantitative protein analysis using mass spectrometry.
Amide Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS) for Epitope Mapping
Yoshi Hamuro, Ph.D., Director, Analysis, ExSAR
Epitope mapping is a critical step for antibody development for scientific, IP, and regulatory reasons. Amide hydrogen/deuterium exchange coupled with proteolysis and mass spectrometry (HDX-MS) can identify a discontinuous conformational epitope in medium-resolution. It is a widely-applicable analytical technique that can address obstacles common to alternative techniques. Furthermore, combining with computational docking, this method can yield high resolution models of antibody-antigen complexes, effectively predicting the actual contact residues on the antigen as verified by co-crystal analysis.
Biography: Dr. Hamuro is an expert of hydrogen/deuterium (H/D) exchange analysis of proteins. He was instrumental in the development of modern H/D exchange technology at the UCSD. He has been leading the efforts on the further improvement of H/D exchange technology at ExSAR and is in charge of developing ExSAR's research strategy for application of H/D exchange technology to drug discovery.
N-linked Glycosylation on Non-consensus Protein Motifs
Alain Balland, Ph.D., Scientific Director, Analytical & Formulation Sciences, Amgen
We have reported through detailed analyses of recombinant antibodies that N-linked glycosylation can be present on asparagine residues that do not adhere to the consensus motif NX(S/T) described in the literature. We will show how a strategy based on lectin enrichment of glycosylated IgGs, followed by endo F2 treatment and peptide mapping with CID-MS2 / ETD or CID-MS3 analysis can help locate several other examples of non-consensus N-linked glycosylation sites. The results point to a common theme around the modified residue, supporting the concept of a novel protein motif for N-glycosylation.
About the Conference:
PEGS is the essential antibody and protein engineering event of the year, where what you learn can be directly applied to your work. The event provides 1,200+ participants to network with their peers and learn about unpublished science in the session rooms. Delegates engage in problem solving breakout discussions and gain insight from over 60 exhibitors, and view the 155 posters in the exhibit hall.