Biophysical Analysis for Bioprocessing
CSS
Post-con
Document Title
Agenda
Wednesday | Thursday
8:00
Registration and Networking Coffee
8:40
Chairperson's Opening Remarks
Steven A. Cohen, Ph.D., RDE Life Sciences Director, Waters Corporation
Steven A. Cohen, Ph.D., RDE Life Sciences Director, Waters Corporation
Keynote Presentation
8:45
Determination of Reversible Self-Association at High Protein Concentration and its Impact on Viscosity
Reversible protein self-association has been linked to high viscosity of a monoclonal antibody (MAb) high concentration formulation (Liu et al., J. Pharm. Sci. 2005). We have used rheological measurements, static light scattering, and preparative analytical sedimentation to investigate the relationship of protein-protein self-association to viscosity in several closely related IgG1 MAbs at concentrations >100 mg/mL A series of charge swap mutants were also studied to evaluate contribution of amino acid residues to self-association and viscosity, and the initial results will be discussed.
Steven J. Shire, Ph.D., Staff Scientist and Group Leader, Late Stage Pharmaceutical and Process Development, Genentech, Inc.
Reversible protein self-association has been linked to high viscosity of a monoclonal antibody (MAb) high concentration formulation (Liu et al., J. Pharm. Sci. 2005). We have used rheological measurements, static light scattering, and preparative analytical sedimentation to investigate the relationship of protein-protein self-association to viscosity in several closely related IgG1 MAbs at concentrations >100 mg/mL A series of charge swap mutants were also studied to evaluate contribution of amino acid residues to self-association and viscosity, and the initial results will be discussed.
Steven J. Shire, Ph.D., Staff Scientist and Group Leader, Late Stage Pharmaceutical and Process Development, Genentech, Inc.
Biophysical Analysis in Early and Late Development: Investigating Protein Structures
9:30
Case
Study
Differential Scanning Calorimetry Studies of the Structure and Function of AbataceptStudy
Abatacept (CTLA4Ig, Orencia®) is a fusion protein that has been approved for the treatment of rheumatoid arthritis. The DSC profile of Abatacept shows two transitions which have melting temperatures of ~58oC and ~83oC. This presentation will describe experiments that identify the regions in the molecule that correspond to the transitions and the correlation of DSC profile with the bioactivity and ligand binding of Abatacept.
Satish K. Mallya, Ph.D., Senior Research Investigator, Biologics Process and Product Development, Bristol-Myers Squibb
10:00
Qualification of Biophysical Methods for Evaluating Secondary and Tertiary Structure of Proteins and use with Analytical Comparability Assessments
During the development of biopharmaceuticals, process changes are often made to fit manufacturing scale, increased yield and the implementation of new technology. A comprehensive analytical comparability assessment is an important part of ensuring product quality after process changes. Biophysical methods are powerful tools to evaluate potential structural change of protein therapeutics. Qualifying biophysical methods adds compliance and assurance to product comparability assessments.
Yen-Huei Lin, Ph.D., Senior Scientist, Drug Product Sciences, Human Genome Sciences, Inc.
During the development of biopharmaceuticals, process changes are often made to fit manufacturing scale, increased yield and the implementation of new technology. A comprehensive analytical comparability assessment is an important part of ensuring product quality after process changes. Biophysical methods are powerful tools to evaluate potential structural change of protein therapeutics. Qualifying biophysical methods adds compliance and assurance to product comparability assessments.
Yen-Huei Lin, Ph.D., Senior Scientist, Drug Product Sciences, Human Genome Sciences, Inc.
10:30
Networking Refreshment Break with Exhibit & Poster Viewing
11:00
Molecular Dynamics Simulations and Biophysical Techniques for Characterization of Protein Dynamics
Antibodies have three major domains in which the Fc domain is connected to the Fab domain via a single peptide linker (hinge). It has been demonstrated previously that the hinge region provides a certain degree of flexibility to antibodies. We are using time-resolved fluorescence anisotropy in conjunction with molecular dynamics simulations to map the different motions around the hinge region. These tools allow us to characterize the solution dynamics of antibodies.
Trevor Swartz Ph.D., Scientist, Early Stage Pharmaceutical Development, Genentech, Inc.
Antibodies have three major domains in which the Fc domain is connected to the Fab domain via a single peptide linker (hinge). It has been demonstrated previously that the hinge region provides a certain degree of flexibility to antibodies. We are using time-resolved fluorescence anisotropy in conjunction with molecular dynamics simulations to map the different motions around the hinge region. These tools allow us to characterize the solution dynamics of antibodies.
Trevor Swartz Ph.D., Scientist, Early Stage Pharmaceutical Development, Genentech, Inc.
11:30
Case
Study
Understanding the Solution Behavior and Higher Order Structure of Protein Biopharmaceuticals Using H/DX-MSStudy
Recent advances in hardware and software are leading to the commercialization of H/DX-MS instruments for studying protein structure and structural dynamics. Such instrumentation should be very useful to the biopharmaceutical industry for biophysical comparability studies and drug design. Hence the focus and goal of this talk will be to discuss opportunities and show examples of how this technology can be used to improve the protein biopharmaceutical development process.
Steven Berkowitz, Ph.D., Principal Investigator, Analytical Development, Biogen Idec
12:00
Networking Luncheon and Exhibit & Poster Viewing
1:10
Chairperson's Opening Remarks
Mariana Dimitrova, Ph.D., Senior Scientist, Formulation Sciences Department, MedImmune
Mariana Dimitrova, Ph.D., Senior Scientist, Formulation Sciences Department, MedImmune
Detection and Characterization of Submicron, Subvisible and Visible Particles
Keynote Presentation
1:15
Status of Compendial Assays and Standards Development for Measurement of Subvisible and Visible Particles
The United States Pharmacopeia provides guideline methods and limits for particle content in parenteral products and ophthalmic solutions. Recent USP harmonization with the Japanese and European pharmacopeias has placed the parenteral assays and limits in a common chapter. However, the assays do not adequately address the broad and emergent array of pharmaceutical formulation types and delivery innovations. The presentation reviews the current methods, limits and available standards for the determination of particulate matter content and considers challenges and changes to these guidelines.
D. Scott Aldrich, Principal Consultant, Ultramikro LLC; Member, United States Pharmacopeia Parenteral Products Industrial Expert Committee
The United States Pharmacopeia provides guideline methods and limits for particle content in parenteral products and ophthalmic solutions. Recent USP harmonization with the Japanese and European pharmacopeias has placed the parenteral assays and limits in a common chapter. However, the assays do not adequately address the broad and emergent array of pharmaceutical formulation types and delivery innovations. The presentation reviews the current methods, limits and available standards for the determination of particulate matter content and considers challenges and changes to these guidelines.
D. Scott Aldrich, Principal Consultant, Ultramikro LLC; Member, United States Pharmacopeia Parenteral Products Industrial Expert Committee
2:00
Applications of Submicron and Subvisible Particle Analysis in the Development of Protein Therapeutics
This abstract was not available at the time of printing the brochure.
Mariana Dimitrova, Ph.D., Senior Scientist, Formulation Sciences Department, MedImmune
This abstract was not available at the time of printing the brochure.
Mariana Dimitrova, Ph.D., Senior Scientist, Formulation Sciences Department, MedImmune
2:30
Considerations in the Development of NIST Standards for Protein Particulates
NIST reference materials provide a path for validation of methodology and/or instrument performance. Existing NIST reference materials for particle size and density poorly duplicate the properties of protein particulates. We report preliminary results on measurement issues for characterization of protein particulates, including the use of surrogates as reference materials, applicability to multiple detection methods, and the need for standardized methods.
Dean Ripple, Ph.D., Group Leader, Thermometry Group, National Institute of Standards and Technology
NIST reference materials provide a path for validation of methodology and/or instrument performance. Existing NIST reference materials for particle size and density poorly duplicate the properties of protein particulates. We report preliminary results on measurement issues for characterization of protein particulates, including the use of surrogates as reference materials, applicability to multiple detection methods, and the need for standardized methods.
Dean Ripple, Ph.D., Group Leader, Thermometry Group, National Institute of Standards and Technology
3:00
Networking Refreshment Break with Exhibit & Poster Viewing
3:30
Survey of Current Understandings of the Role of Subvisible Particles in Aggregation and Immunogenicity
Little is known about the exact mechanisms through which aggregates of therapeutic products induce an immune response. We investigate the ability of homogeneous and heterogeneous aggregates of recombinant growth hormone to provoke immune responses in mice. We find that subvisible protein particles induce T-cell dependent immune responses when administered subcutaneously. Furthermore, we find that the use of high hydrostatic pressure to reduce aggregate levels can reduce and even eliminate immunogenicity.
Amber Fradkin, Ph.D., Postdoctoral Fellow, University of Colorado at Boulder
Little is known about the exact mechanisms through which aggregates of therapeutic products induce an immune response. We investigate the ability of homogeneous and heterogeneous aggregates of recombinant growth hormone to provoke immune responses in mice. We find that subvisible protein particles induce T-cell dependent immune responses when administered subcutaneously. Furthermore, we find that the use of high hydrostatic pressure to reduce aggregate levels can reduce and even eliminate immunogenicity.
Amber Fradkin, Ph.D., Postdoctoral Fellow, University of Colorado at Boulder
4:00
Case
Study
Subvisible Particles in Biopharmaceuticals: Case Studies on Method Development and Links to ImmunogenicityStudy
Sub-visible particles <10 mm are increasingly being monitored during development of biopharmaceuticals. Through case studies, we will describe method development for sub-visible particle characterization, with an emphasis on samples that present challenges to the analytical techniques (e.g. high protein concentration, low conductivity formulation buffer). We will also discuss how the collected data can (or cannot) be correlated with immunogenicity.
Barthélemy Demeule, Ph.D., Scientist, Genentech, Inc.
4:30
Application of FTIR and Microscopy to the Characterization of Subvisible and Visible Particles in Therapeutic Proteins
Evaluation of the nature of subvisible and visible particles is important to ensure successful process development and drug product characterization. The purpose of this study was to evaluate Electronic FTIR Microscopy for identification of subvisible and visible particles in therapeutic proteins. Correlation with other analytical methodologies such as SDS-PAGE and Western Blot will also be presented.
Jinping Liu, Ph.D., Principal Scientist, Bristol-Myers Squibb
Evaluation of the nature of subvisible and visible particles is important to ensure successful process development and drug product characterization. The purpose of this study was to evaluate Electronic FTIR Microscopy for identification of subvisible and visible particles in therapeutic proteins. Correlation with other analytical methodologies such as SDS-PAGE and Western Blot will also be presented.
Jinping Liu, Ph.D., Principal Scientist, Bristol-Myers Squibb
5:00
Networking with Exhibit & Poster Viewing
Wednesday | Thursday
Implementing Best Practices in Biophysical Analysis from Research through Development
8:10
Registration and Networking Coffee
8:35
Chairperson's Opening Remarks
Pooja Arora, Ph.D., Research Investigator, Bristol-Myers Squibb
Pooja Arora, Ph.D., Research Investigator, Bristol-Myers Squibb
8:40
Applications of Biophysical Analysis at Research Stage: The Biophysical Challenge of Measuring Affinities of Tight Antibody/Antigen Complexes
Development of therapeutic MAbs requires rigorous measurements of kinetic and thermodynamic binding properties of antibody/antigen complexes for candidate optimization and clinical dosing strategies. Surface plasmon resonance (SPR) and kinetic exclusion assays (KinExA) can reliably measure single-digit picomolar affinities often associated with therapeutic antibodies. Similar kinetic rate constants and affinities were determined with these solid phase (SPR) and solution phase (KinExA) methodologies when experiments were designed correctly, the instrumentation was utilized properly, and data was processed optimally.
Andrew W. Drake, Ph.D., Principal Scientist, Biophysical Chemistry and Research Informatics, Takeda San Francisco, Inc.
Development of therapeutic MAbs requires rigorous measurements of kinetic and thermodynamic binding properties of antibody/antigen complexes for candidate optimization and clinical dosing strategies. Surface plasmon resonance (SPR) and kinetic exclusion assays (KinExA) can reliably measure single-digit picomolar affinities often associated with therapeutic antibodies. Similar kinetic rate constants and affinities were determined with these solid phase (SPR) and solution phase (KinExA) methodologies when experiments were designed correctly, the instrumentation was utilized properly, and data was processed optimally.
Andrew W. Drake, Ph.D., Principal Scientist, Biophysical Chemistry and Research Informatics, Takeda San Francisco, Inc.
9:10
Use of Biophysical Tools During Early Development: Preformulation, Forced Degradation, and Process Development Applications
A critical first step toward product development is thorough characterization of the protein. Biophysical techniques are an essential component of this initial characterization to facilitate an understanding of the conformational, thermal, physical, and biological stability of the API under a variety of process and formulation conditions. We present examples of how such techniques were used to guide the development process.
Atul Saluja, Ph.D., Group Leader / Scientist, Biopharmaceutical Development, KBI Biopharma, Inc.
A critical first step toward product development is thorough characterization of the protein. Biophysical techniques are an essential component of this initial characterization to facilitate an understanding of the conformational, thermal, physical, and biological stability of the API under a variety of process and formulation conditions. We present examples of how such techniques were used to guide the development process.
Atul Saluja, Ph.D., Group Leader / Scientist, Biopharmaceutical Development, KBI Biopharma, Inc.
9:40
Case
Study
Utility of Thermodynamics in Excipient Screening During Formulations DevelopmentStudy
This abstract was not available at the time of printing the brochure.
Eric L. Reese Ph.D., Director, Biotherapeutics Products, GE Healthcare
10:10
Networking Refreshment Break with Exhibit & Poster Viewing
10:40
Case
Study
Analysis of Leachables and Degraded Excipients as a Cause for Protein ModificationsStudy
Leachables or degraded excipients can be the root cause for apparent protein modifications. In this presentation, two example cases will be presented. An out-of-trend peak in a SEC chromatogram, initially suspected to represent aggregation, was identified as an unexpected leachable by headspace gas chromatography. In the second example an increased rate of protein glycation was linked to the presence of glucose. Investigations to identify the origin of the glucose will be described.
Helmut Schneider, Ph.D., Senior Scientist, Analytical Sciences, Human Genome Sciences, Inc.
Biophysical Analysis in Formulation Development
11:10
Assessing the Physicochemical Properties of Biopharmaceutical Drug Candidates and Formulations using Biophysical Methods
Often drug candidates in discovery and development exhibit undesirable chemical and physical instability upon storage, processing, and delivery. This presentation will describe examples of phase appropriate yet insightful contributions from biophysical analyses in the assessment of a preformulation molecule, the evaluation of a development stage protein formulation, and to study the impact of process induced change to a formulation component.
Derrick Katayama, Ph.D., Senior Development Scientist, Pharmaceutical Sciences, Amylin Pharmaceuticals
Often drug candidates in discovery and development exhibit undesirable chemical and physical instability upon storage, processing, and delivery. This presentation will describe examples of phase appropriate yet insightful contributions from biophysical analyses in the assessment of a preformulation molecule, the evaluation of a development stage protein formulation, and to study the impact of process induced change to a formulation component.
Derrick Katayama, Ph.D., Senior Development Scientist, Pharmaceutical Sciences, Amylin Pharmaceuticals
11:40
Case
Study
Characterizing Protein-Protein Interactions at Moderate and High ConcentrationStudy
Therapeutic proteins are often formulated at relatively high concentrations. Intermolecular interactions that hold the potential for causing aggregation, high visocosity, and other deleterious effects may not be evident in typical measurements at moderate concentrations of a few g/L or less. We demonstrate the application of Composition-Gradient Static Light Scattering to a protein system that behaves quite differently in different formulations, in order to tease out the oligomeric stoichiometry and binding affinity as well as the magnitude of non-specific interactions at play.
Daniel Some, Ph.D., Principal Scientist, Wyatt Technology Corp.
12:10
Networking Luncheon and Exhibit & Poster Viewing
1:35
Chairperson's Opening Remarks
Prathima Acharya, Ph.D., Technical Director and Managing Member, Biosonata Consulting, LLC
Prathima Acharya, Ph.D., Technical Director and Managing Member, Biosonata Consulting, LLC
Biophysical Methods for Comparability Analysis
1:40
Characterization of Antibody-Receptor Interactions by Atomic Force Microscopy
The inter-molecular binding strength of ligand-receptor pairs can be pivotal for many biological processes such as receptor signaling. In addition to the capability of imaging bio-molecules with sub-molecular resolution, the atomic force microscope is suitable for measuring molecular forces at the single molecule level in a liquid environment. In this talk we will show how it is possible to analyze molecular interactions between proteins and ligands, and how it complements data obtained by surface plasmon resonance and bioassay.
Linda Obenauer-Kutner, Senior Research Scientist, Bristol-Myers Squibb
The inter-molecular binding strength of ligand-receptor pairs can be pivotal for many biological processes such as receptor signaling. In addition to the capability of imaging bio-molecules with sub-molecular resolution, the atomic force microscope is suitable for measuring molecular forces at the single molecule level in a liquid environment. In this talk we will show how it is possible to analyze molecular interactions between proteins and ligands, and how it complements data obtained by surface plasmon resonance and bioassay.
Linda Obenauer-Kutner, Senior Research Scientist, Bristol-Myers Squibb
2:10
Improving the Comparability of Protein Circular Dichroism Measurements
Two recent international comparison studies of protein circular dichroism measurements (CD) reveal significant differences between CD spectra from different laboratories. Our talk will highlight the studies and describe the issues raised. Our efforts to compare and correct CD spectra using separate measurements of left and right circularly polarized spectra, rather than artifact standards, will also be described.
Curtis W. Meuse, Ph.D., Research Chemist, National Institute of Standards and Technology (NIST)
Two recent international comparison studies of protein circular dichroism measurements (CD) reveal significant differences between CD spectra from different laboratories. Our talk will highlight the studies and describe the issues raised. Our efforts to compare and correct CD spectra using separate measurements of left and right circularly polarized spectra, rather than artifact standards, will also be described.
Curtis W. Meuse, Ph.D., Research Chemist, National Institute of Standards and Technology (NIST)
2:40
Evaluation of Mechanical Stress Techniques for use in Therapeutic Protein Characterization Studies
This abstract was not available at the time of printing the brochure.
Gregory Walsh, Process Analytical Scientist, Technology Development, Genzyme Corporation
This abstract was not available at the time of printing the brochure.
Gregory Walsh, Process Analytical Scientist, Technology Development, Genzyme Corporation
3:10
Networking Refreshment Break with Exhibit & Poster Viewing
Problem Solving with Multiple Biophysical Methods
3:40
Case
Study
Quantifying Biomolecular Interactions via a Novel Composition-Gradient Static Light Scattering (CG-SLS) Biophysical Method as a Complimentary and Orthogonal Technique to Surface Plasmon Resonance (SPR) TechnologyStudy
This presentation provides antibody-antigen binding data from Composition Gradient Static Light Scattering (CG-SLS) compared to SPR analysis. In one case study, additional information obtained from CG-SLS not only helped determine the unexpected dimeric form of the receptor, but also revealed the true stoichiometric ratio of the antibody-receptor interaction. In another example, the lack of a comprehensive understanding of the structural biology of the antigen-antibody interaction prevented a definitive assignment of the proper binding model in the Biacore® analysis.
Jihong Yang, Ph.D., Scientist, Bioanalytical Research and Development, Genentech, Inc.
4:10
Use of Analytical Ultracentrifugation (AUC) to Characterize Low Molecular Weight Species in Drug Substance Produced at a CMO
A recombinant protein drug substance produced at a CMO was found to have a low molecular weight (LMV) shoulder peak that eluted behind the size exclusion chromatography (SEC) main peak. Analytical ultracentrifugation showed that the LMW species sediments at 3.1S. An assignment of this species by AUC was based on a comparison of this experimental value to that computed using the software HYDROPRO that generates a hydrodynamic bead model from the protein crystal structure.
Nina Xiao, Ph.D., Research Associate, Late Stage Pharmaceutical & Device Development, Genentech, Inc.
A recombinant protein drug substance produced at a CMO was found to have a low molecular weight (LMV) shoulder peak that eluted behind the size exclusion chromatography (SEC) main peak. Analytical ultracentrifugation showed that the LMW species sediments at 3.1S. An assignment of this species by AUC was based on a comparison of this experimental value to that computed using the software HYDROPRO that generates a hydrodynamic bead model from the protein crystal structure.
Nina Xiao, Ph.D., Research Associate, Late Stage Pharmaceutical & Device Development, Genentech, Inc.
4:40
End of Conference
Bottom menu
Copyright IBC Life Sciences, a part of Informa Life Sciences Group