Biophysical & Biochemical Characterization of Biotherapeutics


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4:00 - 6:00 pm Main Conference Registration



7:00 am Registration and Morning Coffee


Latest Tools for Biopharmaceutical Characterization 

8:30 Organizer’s and Chairperson’s Opening Remarks

Micah Lieberman, Executive Director, Conferences, Cambridge Healthtech Institute (CHI)

Jennifer Nemeth Jennifer Nemeth, Ph.D., Principal Research Scientist, Biologics Mass Spectrometry & Allied Technologies, Centocor R&D, Inc.




Introductory Presentation: Overview of Current Analytical Technologies in the Biopharmaceutical Characterization Field, and Where Changes are Required to Address Unmet Needs 

Jennifer Nemeth Jennifer Nemeth, Ph.D., Principal Research Scientist, Biologics Mass Spectrometry & Allied Technologies, Centocor R&D, Inc. 

There are a wide range of analytical tools for the assessment and characterization of biopharmaceuticals. Generally, a set of tools and processes are established, and the path to making changes down the line can be difficult, even in a discovery environment. Obstacles to change are not necessarily related to SOPS and regulations, but to lack of development time and hesitancy to “do something different.” Here, an examination of advances in standard technologies, or a new look at little-used ones, will be provided, and proposals on how these technologies might enhance the biopharmaceutical discovery/development process will be presented. 

9:10 Characterization of Heterogeneous Proteins by Ion/Ion Chemistry Coupled with Ion Mobility Mass Spectrometry

Paul SchnierPaul Schnier, Ph.D., Principal Scientist, Molecular Structure, Amgen, Inc.

Electrospray ionization mass spectrometric (ESI-MS) analysis of heterogeneous proteins often results in complex, unresolved spectra. Here we demonstrate how ion-ion chemistry can be used to enable the analysis of heterogeneous proteins by mass spectrometry. We demonstrate how anions generated with a home built glow discharge source can be used to manipulate the charge states of ESI generated ions, greatly reducing the spectral congestion typically observed for polydisperse molecules. Additionally, we demonstrate how ion-ion chemistry can be used to selectively charge strip protonated protein ions to simplify ESI mass spectra of proteins which form extensive non-specific metal adducts.

9:40 Novel Chemical Pathways of Protein Degradation: Characterization of Products and Immunogenicity

Christian Schoneich Christian Schöneich, Ph.D., Professor, Chair, Pharmaceutical Chemistry, University of Kansas

Chemical degradation presents an important problem for the design of stable formulations of therapeutic proteins. Chemical degradation may trigger aggregation and/or fragmentation, and lead to novel epitopes on proteins, all of which may contribute to immunogenicity. Several pathways of chemical degradation are known such as hydrolytic, oxidative and photolytic pathways. This presentation will focus especially on oxidative and photolytic pathways with focus on novel reactions of peptide and protein cysteine, cystine, and methionine in both solution and the solid state, and on the characterization of the products of these pathways.

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

11:10 Relative Sensitivity of Common Biophysical Methods and Conventional Chromatographic and Functional Assays to Changes in Protein Higher Order Structure

Andrew Kosk Andrew Kosky, Ph.D., Senior Group Leader, Early Stage Pharmaceutical Development, Genentech

We have compared commonly used biophysical methods (e.g. circular dichroism and Fourier-transform infrared spectroscopy) and conventional chromatographic and functional assays to determine which types of methods are most sensitive to higher order structural changes in proteins. Our results demonstrate that commonly used biophysical techniques are often less sensitive than conventional purity and potency assays to the types of structural changes that impact protein function (in vitro) and overall therapeutic protein product quality.

11:40 A Platform Independent Analysis System for the Characterization of Chemical Liability in Biotherapeutics

Steven Pomerantz Steven Pomerantz, Ph.D., Senior Research Scientist, Centocor R&D, Inc.

After winnowing the biochemical diversity through various characterizations and assays, a biotherapeutic development program may still be faced with several candidate molecules with equivalent affinity or bioactivity. One criterion for the selection of a lead candidate molecule is its developability, a suite of requirements to maximize the desirable chemical and biophysical attributes, and minimize potential chemical liabilities, such as post-translational modification (PTM). We have developed an MS-based system coupled with back-end quantitation and identification software for the rapid analysis of defined PTMs to enable screening of larger panels of candidate molecules to ensure maximum information availability for selection of the therapeutic lead.

12:10 pm On-line SAW-Bioaffinity-Mass Spectrometry: New Bioanalytical Application in Detection, Structure Determination and Quantification of Biomolecular Protein-Ligand Interactions from Biological Material

Michael Przybylski Michael Przybylski, Chair, Laboratory of Analytical Chemistry and Biopolymer Structure Analysis, University of Konstanz, Germany

In recent years, bioaffinity analysis using biosensors such as surface plasmon resonance has become an established technique for the detection and quantification of biomolecular interactions; however, a severe limitation of biosensors is their lack of providing structure information of affinity-bound biopolymer ligands. Bioaffinity- mass spectrometry is a new, combined analytical approach for the detection, quantification, and structure determination of affinity-bound ligands. We have developed an online combination of a surface acoustic wave (SAW) biosensor with electrospray ionization mass spectrometry (SAW-ESI-MS) that enables the direct structure determination and quantification of affinity-bound ligands dissociated from a protein-ligand complex on a gold chip. An interface for the coupling of SAW-biosensor chip and ESI- MS provides a sample concentration and in-situ desalting step for the MS analysis of the ligand eluate solution. First applications of the online SAW-MS combination with chip-immobilized antibody and polypeptide ligands show broad bioanalytical application to the simultaneous, label-free structure determination and quantification of biopolymer-ligand interactions, as diverse as antigen-antibody and lectincarbohydrate complexes. Dissociation constants (KD) are determined in the milli- to nanomolar range directly from biological material, such as cell lysate and brain homogenate.

12:40 Luncheon Presentations (Sponsorship Opportunities Available) or Lunch on Your Own

1:40 Break


Biophysical Characterization of Therapeutic Proteins 

2:00 Chairperson’s Remarks

Andrew Kosk Andrew Kosky, Ph.D., Senior Group Leader, Early Stage Pharmaceutical Development, Genentech



2:05 Interpretation and Misinterpretation of Biophysical Analysis in Characterization, Formulation and Process Development of Protein Therapeutics, and its Application in High Throughput Formulation Screening

Haripada MaityHaripada Maity, Ph.D., Principal Scientist, Formulation Development, ImClone Systems, a wholly owned subsidiary of Eli Lilly & Co.

Protein structure is stabilized enthalpically and enthalpy-entropy compensation makes protein marginally stable. Biophysical analysis plays a unique role in characterizing higher-order structure and evaluating the stability of complex protein molecules. This presentation will discuss a variety of case studies that involve (i) prediction of optimized formulation based on structure and conformational stability and caveats in the use of accelerated stability data in formulation selection, (ii) cautionary notes for the analysis of accelerated stability samples, (iii) understanding different thermal and thermodynamic stability parameters, (iv) steady-state and kinetic analysis in the optimization of protein stability in low pH for process development, (v) sensitive biophysical methods for comparability assessment, and (vi) challenges in the use of biophysical techniques in high throughput formulation screening.

2:35 Challenges in Testing and Characterization of Bionanotherapeutics

Nanda Subbarao Nanda Subbarao, Ph.D., Senior Consultant, Analytical CMC, Biologics Consulting Group

The physiological action of Bionanotherapeutics depends strongly on the size and morphology of the nanoparticles; therefore, their testing and characterization must include methods which address these parameters. Characteristics of the nanoparticle matrix and its interaction with the drug has to be studied, in addition to analytical methods commonly required for all biotherapeutics. Methods useful for characterization, lot release and stability studies on bionanotherapeutics and common ways in which nanoparticles interfere with these assays will be presented.

3:05 Refreshment Break in the Exhibit Hall with Poster Viewing

3:45 Composition-Dependent Properties of Monoclonal Antibody Formulations

Jonas Fast Jonas Fast, Ph.D., Formulation Scientist, Pharmaceutical and Analytical R&D Biologics, F. Hoffmann-La Roche

During development of a monoclonal antibody formulation, significant composition-dependent physical bulk properties and chemical degradations were observed. The driving forces of these observations were investigated in several systems.


4:15 Correlation between the Differential Scanning Profile, Binding Activity and Bioactivity of Abatacept

Satish Mallya Satish Mallya, Ph.D., Senior Research Investigator, Biologics Process and Product Development, Bristol-Myers Squibb

Abatacept (CTLA4Ig) is a fusion protein that has been approved for the treatment of rheumatoid arthritis. The Differential Scaning Calorimetry (DSC) profile of abatacept shows two major transitions with melting temperatures of 58C and 83C. This presentation will discuss the correlation between the transitions, ligand binding activity and bioactivity of abatacept.



4:45 Problem Solving Breakout Sessions 

Concurrent Problem Solving Breakouts are interactive sessions hosted by a moderator to discuss a topic in depth. They are open to all attendees, sponsors, exhibitors, and speakers and provide a forum for discussing key issues and meeting potential partners. Please pick a topic of your choice and join in. 

TABLE 14:  Disulfide Bond Mapping Using Mass Spectrometry 

Moderator:  Dariusz Janecki, Ph.D., Research Scientist, Biologics Mass Spectometry, Centocor R&D, Inc. 

• Disulfide bond mapping of monoclonal antibodies
• Highly knotted, cysteine rich proteins, use of alternative enzymes for digests
• Analysis methodologies

TABLE 15:  Biophysical Analysis in the Characterization and Development of Protein Pharmaceuticals 

Moderator:  Haripada Maity, Ph.D., Principal Scientist, Formulation Development, ImClone Systems 

• Sensitive biophysical methods for comparability assessment of protein pharmaceuticals.
• Sensitive technique to optimize stability of protein in low pH and its application in process development.
• Linkage analysis of stability parameters obtained from DSC and optical spectroscopic techniques.

TABLE 16:  Evaluation of Comparability of Protein Therapeutics Due to Scale Up, Change in Production Facilities, Fermentation, Purification and Formulation 

Moderator:  Brent Kendrick, Ph.D., Director, Analytical Sciences, Amgen 

• How to balance statistical equivalence criteria with practical / scientifically derived assessment of comparability
• Forced degradation studies for comparability assessment (how to set up, when to use, which sample types apply, etc)
• Biochemical and Biophysical methods for comparability assessment

TABLE 17:   Detecting and Characterizing of Soluble and Insoluble Protein Aggregates: Novel Methods; Complementary/Orthogonal Analytical Tools 

Moderator:  Nanda Subbarao, Ph.D., Senior Consultant, Analytical CMC, Biologics Consulting Group
Moderator:  Devendra (Davy) S. Kalonia, Ph.D., Professor of Pharmaceutics, Department of Pharmaceutical Sciences University of Connecticut 

• Aggregate nomenclature and types of aggregates
• Size characterization
• Structural Characterization

TABLE 18:  Analytical Characterization Requirement for Supporting FDA Filings   

Moderator:  Jennifer Nemeth, Ph.D., Principal Research Scientist, Biologics Mass Spectrometry & Allied Technologies, Centocor R&D, Inc. 

• What changes in analytical requirements are being observed for filings, compared to the past
• What new technologies are having an impact for successful filings
• What analytical challenges exist today which are impacting BLA filings

TABLE 19:  Evaluating Comparability and Biosimilarity: Preclinical and Clinical Considerations 

Moderator:  Shefali Kakar, Ph.D., Clinical Pharmacology, Oncology Business Unit, Novartis 

• Is comparability different from biosimilarity?
• Extent of preclinical data required?
• Preclinical immunogenicity?
• Clinical data? How much and why?

TABLE 20:  Developing Processes to Manufacture Unstable Proteins into a Stable Dosage Form 

Moderator:  Charlie Hitscherich Jr., Ph.D., Senior Development Engineer, Sterile & Liquids Commercialization, Biogen Idec, Inc. 

• Evaluating compatibility of your drug product with the desired container closure
• Overcoming process induced degregation issues to develop a viable commercial manufacturing process
• Challenges in manufacturing high concentration protein doseage forms
• Key considerations to incorporate into your development plans if you are considering adding an autoinjector

5:45 - 6:45 Reception in the Exhibit Hall with Poster Viewing

Recommended Short Courses 

SC5 – Biolological Mass Spectrometric Applications for Drug Discovery and Product Development 

SC9 – Characterization Techniques for Protein Therapeutics – Orthogonal vs. Complimentary 

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