2013 Archived Content
August 19-20, 2013
Cambridge Healthtech Institute’s Second Annual
Higher-Order Protein Structure: Characterization and Prediction
Day 1 | Day 2 | Short Courses | Download Brochure
Tuesday, August 20
8:00 am Morning Coffee
8:25 Chairperson’s Remarks
Marina Kirkitadze, Ph.D., MBA, Deputy Director, Head, Biophysics and Conformation Unit, Biochemistry Platform Analytical R&D North America, Sanofi Pasteur Ltd.
8:30 FEATURED PRESENTATION:
Protein Clustering and Viscosity
Thomas Laue, Ph.D., Professor, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire - Biography
High concentration protein solutions can exhibit excessively high viscosity. The underlying cause of viscosity is momentum transfer, and the asymmetry of solution components is of great importance in determining momentum transfer. This talk will focus on how proximity energies can contribute to the formation of loose, asymmetric protein clusters that will result in high viscosity.
9:00 Examining Principles of Conformational Switching Using Engineered Proteins of Similar Sequence
Philip N. Bryan, Ph.D., Institute for Bioscience and Biotechnology Research and Department of Bioengineering, University of Maryland - Biography
Certain proteins can adopt different tertiary and quaternary structures as a result of minor perturbation. This fact has implications in a number of important areas including computational and structural biology, protein evolution, human disease, and the design of protein pharmaceuticals. We are examining basic principles sequence-structure relationships using designed proteins that are have different folds but are very similar in sequence. In this simplified sequence space we explore mutational paths from one fold and function into another.
9:30 The Structural Characterization of IgG Particles Formed as a Consequence of Surface Interaction
Tatiana Perevozchikova, Ph.D, Researcher, nSoft Consortium, University of Delaware/ National Institute of Standards and Technology - Biography
Destructive association of biotherapeutics with various surfaces is one of the proposed mechanisms for induction of protein aggregation. Here we demonstrate how our preliminary findings on the interfacial properties of IgGs correlate with the structural information on particles formed upon desorption. Using multiple techniques — neutron reflectivity, CD, MFI and fluorescence — we reveal the consequence of protein adsorption/desorption on conformational dynamics of particle formation.
10:00 Sponsored Presentation (Opportunity Available)
10:15 Coffee Break in the Exhibit Hall with Poster Viewing
11:00 Analysis of the Size, Shape, and Composition of Higher-Order Protein Structures
Lumelle A. Schneeweis, Ph.D., Senior Investigator, Protein Science & Structure, Bristol-Myers Squibb Company - Biography
Characterization of the size, shape, and composition of higher order protein structures often requires a combination of analytical methods for studying the oligomeric state of proteins. Static light scattering (SEC-MALS, AFFF-MALS), sedimentation velocity and equilibrium analytical ultracentrifugation, and less routine methods such as solution atomic force microscopy (AFM) have been used to study the oligomerization of protein conjugates and assemblies.
11:30 Hydrogen / Deuterium Exchange Mass Spectrometry: A Valuable Tool for Protein Higher-Order Structure Characterization
Roxana E. Iacob, Ph.D., Research Assistant Professor, Department of Chemistry and Chemical Biology, Barnett Institute, Northeastern University - Biography
Mass spectrometry (MS) has an important role to play in structural characterization of proteins. Hydrogen exchange (HX) MS has recently become more widely used in the biopharmaceutical industry for protein analysis. Recent advances that make the method accessible to all will be described along with examples of the application of HX MS to matters in the industry.
12:00 pm NMR Methods to Monitor Protein Aggregation and Monomer-Aggregate Interaction Kinetics at Atomic Resolution
Nicolas Fawzi, Ph.D., Assistant Professor, Department of Molecular Pharmacology, Physiology and Biotechnology, Brown University - Biography
High-resolution NMR techniques complement other tools by offering atomic resolution characterization of the aggregation of biopharmaceuticals directly in solution. Rapid, non-destructive measurements allow both the quantitation of soluble protein over time and the determination of the interaction kinetics of soluble species with aggregated material. New NMR methods can characterize the atom-by-atom structure and dynamics of these monomer-aggregate complexes.
12:30 Sponsored Luncheon Presentation (Opportunity Available) or Lunch on Your Own
1:55 Chairperson’s Remarks
Daniel Some, Ph.D., Principal Scientist, Wyatt Technology Corporation
2:00 Higher-Order Structure Analysis for the Development and Trouble-Shooting of Biopharmaceuticals
Gabriella Leo, Ph.D., Associate Researcher, Structural and Biophysical Characterization Laboratory, Analytical Development Biotech Products, Merck Serono (Italy) - Biography
A detailed knowledge of the protein structure is a prerequisite for the development of biopharmaceuticals. The conformation of a protein determines its function and is largely defined through its primary structure, although it can also be significantly influenced by post-translational modifications (PTMs). In this talk, two different case studies will be presented in which the state-of-the-art analysis of higher-order structure is fundamental. The first one deals with the modification of specific amino acids resulting in a bioactivity-diminishing conformational change. The second one deals with an investigation on conformers of a therapeutical antibody initially detected by analytical ultracentrifugation.
2:30 Position Specific Effects of Chemical Composition on Protein Stability
Andria L. Skinner, Ph.D., Scientist, Formulation Development, Regeneron Pharmaceuticals, Inc.
A protein’s stability is determined by its chemical composition, environment and solution conditions. A study of how proteins are affected at the molecular level by intrinsic and extrinsic factors will be presented.
3:00 Application of High-Resolution NMR in the Characterization of Protein Therapeutics
John P. Marino, Ph.D., Leader, Biomolecular Structure & Function Group, National Institute of Standards and Technology - Biography
High-resolution NMR methods can yield spectral ‘fingerprints’ related to the structure of the bioactive form(s) of protein therapeutics at atomic resolution. This presentation will focus on methods for obtaining NMR spectral ‘fingerprints’ of protein therapeutics and how these ‘fingerprints’ can be used to establish consistency in drug manufacturing and for comparing a biosimilar to an innovator reference product.
3:30 Refreshment Break in the Exhibit Hall with Poster Viewing
4:15 Analytical Ultracentrifugation (AUC) as an Orthogonal Method to Characterize Protein Aggregation and Aligning AUC with other Aggregate-Sensitive Techniques
George Svitel, Ph.D., Senior Scientist, Process & Product Development, Amgen, Inc. - Biography
AUC is widely used orthogonal technique to analyze aggregation and regulatory agencies are increasingly requesting AUC data. AUC is able to detect aggregates not reported or under-reported by other techniques. Data show differences in aggregate levels in long-storage, in-process and accelerated degradation samples detected by various techniques. Combining AUC with other techniques opens a path to better understanding of aggregation pathways and mechanisms.
4:45 Orthogonal Particle Sizing Methods in Biopharmaceutical Development
Christopher Mensch, Scientist, Vaccine Drug Product Development, Merck Research Laboratories - Biography
Measuring size distributions of heterogeneous and dynamically changing particles in therapeutic protein and vaccine formulations is a challenging task that requires the use of multiple approaches. Examples of the application of various sizing methods, including micro-flow imaging (MFI), Nanosight, laser diffraction, flow cytometry, atomic force microscopy, dynamic light scattering and colloidal sedimentation velocity (LumiSizer) will be discussed.
5:15 End of Day & Registration for Dinner Short Courses
6:00 Dinner Short Courses*
*Separate registration required
Day 1 | Day 2 | Short Courses | Download Brochure