PXC2


Biophysical analysis is now playing a significant role in the research and early development for a new generation of complex protein therapeutics. The combination of improved understandings of the structure and function of proteins with advanced, higher resolution analytical methods helps researchers identify and fine-tune candidate molecules before advancing them into development. As protein engineers and analytical scientists increase their reliance on biophysical analysis, they are driving a move to instruments with higher throughput and resolution – and working to quantify analytical results previously used only for qualitative assessments. The Biophysical Analysis meeting at PEGS will bring together an international audience of protein scientists and analytical specialists to explore the latest technologies and techniques used for problem solving in this dynamic field.

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Recommended Pre-Conference Short Course*

SC9: Overcoming the Challenges of Immunogenicity Assessment

*Separate registration required


WEDNESDAY, APRIL 27

7:00 am Registration and Morning Coffee

8:00 Chairperson’s Remarks

Joel Bard, Ph.D., Principal Research Scientist, Global Biotherapeutic Technologies, Pfizer


8:10 KEYNOTE PRESENTATION:

Designing and Evolving Antibodies with Improved Biophysical Properties

Peter M. Tessier, Ph.D., Associate Professor, Chemical & Biological Engineering, Rensselaer Polytechnic Institute

Methods for rapidly generating and optimizing the properties of monoclonal antibodies are important for a wide range of applications. I will discuss our progress in improving methods for designing, evolving and selecting of single- and multidomain antibodies with stability and solubility in addition to high affinity.


AUTOMATION AND COMPUTATIONAL TOOLS IN BIOPHYSICAL ANALYSIS

8:40 In Silico Tools for Predicting PK (Clearance) and Viscosity

Vikas Sharma, Ph.D., Senior Group Leader and Senior Scientist, Late Stage Pharmaceutical Development, Genentech

Delivery of high doses of monoclonal antibodies (mAbs) into the subcutaneous space necessitates that mAb solutions exhibit low viscosity and concomitantly demonstrate normal in vivo clearance to enable less frequent dosing. Herein, novel in silico tools are discussed that provide rapid assessment of atypical behavior with respect to high viscosity and faster in vivo clearance. Strikingly, these behaviors are predicted from simple sequence-derived parameters of hydrophobicity, charge dipole distribution and net charge.

9:10 Database Systems for Consolidation and Analysis of Biotherapeutic Molecular Assessment Results

Joel Bard, Ph.D., Principal Research Scientist, Global Biotherapeutic Technologies, Pfizer

Global Biotherapeutic Technologies at Pfizer uses a number of biophysical and biochemical assays to predict the developability of early stage biotherapeutics. Data from these assays is used to remove sequence liabilities before they can cause problems in production. To improve access to this data and allow analysis across multiple projects, we have recently implemented a database to store these results and provided web applications to simplify its deposition and analysis.

9:40 Understanding Protein Dynamics from Empirical Modeling and Fast Screening

Donald Jacobs, Ph.D., Professor, Physics and Optical Science, University of North Carolina

A critical factor to successful rational drug discovery is the use of representative conformational ensembles to capture the role protein dynamics has on function, kinetics and stability through conformational entropy. To overcome computational challenges in constructing and utilizing conformational ensembles for in silico screening applications, the costly molecular dynamics paradigm is replaced with a fast all-atom empirical interfacial thermodynamic model. The link between protein dynamics and thermodynamics is then elucidated.

10:10 Coffee Break in the Exhibit Hall with Poster Viewing


Analysis of Complex Biophysical Data

10:55 Application of Differential Scanning Calorimetry in Biotherapeutic Comparability

William M. Matousek, Ph.D., Staff Scientist, Protein Biochemistry Preclinical Development, Regeneron Pharmaceuticals, Inc.

Differential Scanning Calorimetry (DSC) is a sensitive technique commonly used to define protein thermal unfolding transitions, assess relative thermal stability, and compare high order structural content. Multiple characteristic unfolding transitions are often observed, making the method amenable for both establishing identity and the estimation of impurities. Thermodynamic properties such as Tm, enthalpy, and heat capacity were evaluated as indicators of structural similarity and purity.

11:25 A Showcase of the Application of Biophysical Analysis in Formulation and Process Development: Interpretation and Misinterpretation of Biophysical Data

Haripada Maity, Ph.D., Research Advisor, Formulation Development, Eli Lilly and Company

Enthalpy-entropy compensation causes protein structure to be marginally stable. Therefore, in order to reduce both physical and chemical degradation of a protein, the primary objective is to maximize both its conformational and colloidal stabilities. This presentation will discuss case studies that evaluate (i) the relationship among apparent solubility, conformational stability and protein-protein interactions, (ii) concentration dependent inverse correlation of aggregation, (iii) heat-induced viral inactivation, and (iv) chemical and cold denaturation.

11:55 Supporting Development of Biologic Drugs with Biopharmaceutical Informatics

Satish Singh, Ph.D., Research Fellow and Group Leader, Pfizer

Proactive elimination or mitigation of product development challenges can assure efficient translation of biologic drug candidates into innovative products. Application of computational tools such as informatics, data analyses, modeling and simulations at the earliest stages of product development (i.e., during molecular screening and discovery) helps ensure selection of well-behaved biologic drug candidates. Application of in silico tools will be illustrated with the objective of addressing chemical degradation and viscosity.

Biametrics12:25 pm Presentation to be Announced 

12:55 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on your Own

1:55 Session Break


Emerging Biophysical Analytical Methods

2:10 Chairperson’s Remarks

Robert Walters, Ph.D., Senior Scientist, Pfizer

2:15 Developing Protein-Polymer Conjugates for Ocular Therapeutics

Whitney Shatz, MS, Senior Scientific Researcher, Genentech

Studies with intact antibodies and antibody fragments have suggested a size dependence of vitreal clearance and ocular tissue distribution. Protein PEGylation offers an attractive avenue for potentially increasing size and decreasing vitreal clearance. However, protein‐polymers are complex molecules and analytical characterization is often challenging. The work here describes the development of a biophysical toolbox that has enabled systematic characterization of a range of potential therapeutic agent-polymer (TAP) conjugates.

2:45 Case Study: Novel Biophysical Approaches to High Concentration Formulation Development

Robert Walters, Ph.D., Senior Scientist, Pfizer

High concentration formulations of therapeutic proteins are desirable as they may enable the delivery of an efficacious dose through subcutaneous injection, which is preferable to intravenous administration. Unfortunately, high concentration formulations often suffer from elevated viscosity that may make subcutaneous injection infeasible. This case study will highlight biophysical and computational approaches to creating novel viscosity reducing agents to facilitate high concentration formulation development.


Smaller, Faster, Deeper: Advanced Light Scattering Tools for Biophysical Characterization

3:15 Smaller, Faster, Deeper: Advanced Light Scattering
Tools for Biophysical Characterization

John Champagne, Senior Applications Scientist, Northeast Regional Manager, Wyatt Technology Corp.

3:45 Refreshment Break in the Exhibit Hall with Poster Viewing


4:45 Problem-Solving Breakout Discussions

Importance of Protein Conformational Ensembles to Drug Discovery

Donald Jacobs, Ph.D., Professor, Physics and Optical Science, University of North Carolina

  • Problems with static structures overemphasizing atomic packing and missing entropic effects
  • The role of structural fluctuations on stability, substrate binding and allosteric response
  • Mitigating hypersensitivity while increasing robustness through conformational ensembles
  • How to computationally generate thermodynamically relevant conformational ensembles
  • How to experimentally cross validate representative conformational ensembles

Characterization of Higher Order Structure for Therapeutic Proteins

Jihong Yang, Ph.D., Senior Scientist, Bioanalytical Sciences, Genentech, Inc.

  • Emerging technologies on HOS characterization
  • Development on in silico predication of HOS
  • Application of HOS characterization in drug development
  • Regulatory requirement on HOS characterization
  • Biosimilar experience

Subvisible Particles and Biotherapeutics

Atanas Koulov, Ph.D., Group Head, Senior Principal Scientist, Pharma Technical Development Europe (Biologics) Analytics, F. Hoffmann-La Roche Ltd , Switzerland

  • Particle characterization methods and their application in development
  • Biological consequences of subvisible particles in biotherapeutics
  • Product-specific limits for subvisible particles – how to set specifications?
 

5:45 Networking Reception in the Exhibit Hall with Poster Viewing

7:00 End of Day

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THURSDAY, APRIL 28

8:00 am Morning Coffee


New Developments in Particle Analysis

8:30 Chairperson’s Remarks

William M. Matousek, Ph.D., Staff Scientist, Protein Biochemistry, Regeneron Pharmaceuticals, Inc.

8:35 Biases between Different Particle Counting Instruments in the 1 µm to 100 µm Range

Dean Ripple, Ph.D., Group Leader, NIST

Particle counting instruments may give particle concentrations that differ by a factor of ten or more, especially for proteinaceous particles. This talk presents experimental data and models that describe these biases for light obscuration, flow imaging, and electrical sensing zone techniques. Prospects and challenges on applying reference materials and instrument models to the correction of these biases will also be discussed.

9:05 The Known Unknowns in Subvisible Particle Characterization – Factors Governing the Analytical Performance of Subvisible Particle Measurement Methods

Atanas Koulov, Ph.D., Director, Analytical Development and QC, Drug Product Services, Lonza, Switzerland

Although the biopharmaceutical community is actively using a number of techniques for subvisible particle characterization, the current knowledge of the analytical performance of these tools is inadequate to support their routine use in the development of biopharmaceuticals. This talk will outline some recent efforts to increase this knowledge by systematic evaluation of the analytical performance of the principal subvisible particle characterization techniques and also will provide analysis of the fundamental factors governing it.

9:35 Sponsored Presentation (Opportunity Available)

10:05 Coffee Break in the Exhibit Hall with Poster Viewing


ANALYSIS OF HIGHER ORDER STRUCTURES FOR FINGERPRINTING AND COMPARABILITY ANALYSIS

11:05 A New Method for Oxidative Mapping of Molecular Interfaces

Michael D. Brenowitz, Ph.D., Professor, Biochemistry, Albert Einstein School of Medicine

New tools are needed for the development of protein therapeutics. Indirect structure mapping (‘footprinting’) can define solvent accessible surface with as fine as single residue resolution. We have developed a new approach, Pyrite Shrink-Wrap Laminate that supports hydroxyl radical generation via Fenton chemistry for quantitative protein oxidation. Our protocol is inexpensive to conduct, parsimonious with precious material, easy to implement with a laboratory pipet, and scalable to high-throughput implementation.

11:35 High Order Structures (HOS) Elucidation Using Orthoganol Technologies with Case Studies

Jihong Yang, Ph.D., Senior Scientist, Bioanalytical Sciences, Genentech

Higher Order Structure (HOS) is important to ensure the biological functions of protein and antibody therapeutics. Sensitive and robust technologies that can accurately reveal HOS information and subtle changes are therefore valuable for fingerprinting and comparability analysis, and for drug product stability and process control. Emerging technologies that can be used to elucidate HOS were evaluated using a mAb therapeutic case study, and results from orthogonal methods will be discussed.

12:05 pm Critical Review of HOS Fingerprinting Methods

Tapan Das, Ph.D., Director, Biologics Molecular and Analytical Development, Bristol-Myers Squibb

Structural characterization is an integral part of biotherapeutic development. However, appropriate scrutiny of structural methods is key to ensure the chosen methods serve the intended purpose – based on sensitivity of a method and phase-appropriateness. This talk is intended to provide a critical review of selected higher order structure fingerprinting methods along with examples for utility of such data.

12:35 End of Biophysical Analysis of Biotherapeutics

5:15 Registration for Dinner Short Courses


Recommended Dinner Short Course*

SC12: Strategic Bioassay Design and Analysis

*Separate registration required


Add these programs to your agenda for a full week of comprehensive coverage!

Characterization of Biotherapeutics

Protein Aggregation and Stability in Biopharmaceuticals



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