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Fourth Annual
Protein Aggregation and Emerging Analytical Tools:
Overcoming Analytic, Formulation, Manufacturing, and Regulatory Challenges
January 24-25, 2013 
 

  

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FRIDAY, JANUARY 25

7:15 am Breakfast Presentation (Sponsorship Opportunity Available) or Morning Coffee

 

Improving Resistance to Aggregation 

8:15 Chairperson’s Remarks

Deirdre Piedmonte, Ph.D., Principal Scientist in Drug Product Development, Process and Product Development (P&PD), Amgen

8:20 General Strategy for the Generation of Human Antibody Reagents with Increased Aggregation Resistance

Romain Rouet, Graduate Student, Garvan Institute of Medical Research

Daniel Christ, Ph.D., Group Leader, Immunology Department, Director, Antibody Development, Garvan Institute of Medical Research

The availability of stable human antibody reagents would be of considerable advantage for research, diagnostic, and therapeutic applications. Unfortunately, antibody VH and VL domains that mediate the interaction with antigen have the propensity to aggregate. We have identified mutations that endow superior biophysical properties (non-aggregating, well-expressed, and heat-refoldable) onto human variable domains. Effects of the mutations are highly positional and independent of CDR diversity. Crystal structures reveal a surprising degree of structural conservation, indicating compatibility with VH∕VL pairing and antigen binding. This allows the retrofitting of existing antibody therapeutics.

8:55 Engineering Antibodies for Improved Pharmaceutical Properties

Justin Caravella, Ph.D., Senior Scientist, Physical Biochemistry, Biogen Idec

Development of antibodies can be complicated by their poor biochemical and biophysical properties, such as a tendency to aggregate. We describe knowledge-based and structure-based approaches to engineer antibodies with improved pharmaceutical properties and discuss examples in which an antibody’s development risks were mitigated. Two different monoclonal antibodies have been engineered to have a more than 10-fold increase in solubility by targeted mutagenesis. Successful approaches include reduction of hydrophobicity in key regions and alteration of the antibody’s distribution of charged residues.

9:30 How Solvent and Excipient Properties Impact Aggregation and Viscosity

Thomas Laue, Ph.D., Professor, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New Hampshire

Aggregation and viscosity are sensitive to pH, salt concentrations and salt types, as well as the addition of various excipients. How do these solution properties impact the protein interactions that lead to aggregation and high viscosity? This talk will provide a guide for how to manipulate solvent properties to minimize protein aggregation and reduce viscosity.

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

11:00 Aggregation of Isolated Single IgG1 CH2 Domains

Dimiter Dimitrov, Ph.D., Senior Investigator, Protein Interaction Group, FNLCR, NCI, NIH

The human immunoglobulin (Ig) constant CH2 domain is a promising scaffold for novel candidate therapeutics. We have shown that the stability of CH2 could be increased by introduction of additional disulfide bond and removal of N-terminal seven amino acids (Gong, et al., JBC, 2009 and 2011). However, we found three major aggregation prone regions in CH2, which could lead to aggregation and constrain the use of CH2 scaffold and Fc-related proteins as candidate therapeutics. Therefore, we extensively characterized the aggregation propensity of isolated wild type CH2 and stabilized variants and used various strategies to improve resistance to aggregations which will be discussed.

 

Analytical Tools and Methods 

11:30 Simultaneous Multiple Sample Light Scattering (SMSLS): A New Tool for Continuous Monitoring of Aggregation in Protein Formulations

Wayne Reed, Ph.D., Professor of Physics, Physics and Engineering Physics, Tulane University

Simultaneous Multiple Sample Light Scattering (SMSLS) monitors the kinetics of protein aggregation for many independent protein formulations under varying conditions of temperature, concentration, additives, etc. This high-throughput method runs non-stop and vastly accelerates formulation characterization. SMSLS results on monoclonal antibody aggregation are shown from a collaboration with Biogen Idec.

Malvern Instruments 12:00 pm Recent Advances in the Integrated Measurements of Size, High Order Structure and Formulation Viscosity of Biopharmaceutical Formulations

E. Neil Lewis, Ph.D., Chief Technology Officer, Malvern Instruments

Co-authored by Kevin Mattison, Ph.D., Principal Scientist, Bioanalytics, Malvern Instruments

The non-destructive determination of numerous physicochemical properties of protein therapeutics is critical for developing optimal formulation conditions to enhance product efficacy, stability and manufacturability. To meet this demand, novel optical technologies are being developed to rapidly measure viscosity, high order protein structure and hydrodynamic size from microliter sample volumes.  In this talk we will describe several of these innovations, and how they are able to provide unique insight into the properties and behaviour of biopharmaceutical formulations.

12:30 Close of Morning Session

12:45 Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own

 

Analytical Tools and Methods (Cont.) 

2:00 Chairperson’s Remarks

Wayne Reed, Ph.D., Professor of Physics, Physics and Engineering Physics, Tulane University 

2:05 A Novel Computational Method to Improve Stability, Biophysical Properties, and Expression Levels of Therapeutic Antibodies

Gunasekaran Kannan, Ph.D., Principal Scientist, Therapeutic Discovery, Antibody Optimization, Amgen

We have developed a novel computational method to improve thermal stability and shelf life of antibodies with reduced aggregation levels. The method has been applied to more than two dozens of antibodies from multiple therapeutic programs over last four years. We were able to achieve significant improvement in thermal stability with consistent improvement in the expression levels. 

2:35 Emerging Mass Spectrometry-Based Methods to Characterize Binding and Aggregation of Protein Therapeutics

Igor Kaltashov, Ph.D., Professor, Bioanalytical Chemistry, University of Massachusetts-Amherst

Despite being a very significant problem in the biopharmaceutical sector, protein aggregation is a phenomenon for which the selection of analytical tools remains very limited. Mass spectrometry (MS) has recently emerged as a technique capable of characterizing protein binding and interaction in great detail. Although most of its current uses focus on ordered association of multi-unit protein drugs to their functional form(s) and/or their interactions with therapeutic targets, significant progress has been made recently in developing MS-based methods to study protein aggregation as well. These developments (along with several examples) will be focus of this presentation.

3:05 How Can Protein Formulation Science Benefit from Emerging Techniques for Subvisible Aggregate Characterization?

Andrea Hawe, Ph.D., Head, Scientific Development, Biopharmaceutical R&D, Coriolis Pharma

Within the talk I will focus on the scientific evaluation of novel techniques for subvisible aggregate analysis (e.g. Archimedes, Coulter Counter) compared to the current techniques (e.g. flow imaging, light obscuration, DLS), in particular on their use for protein formulation. Benefits and drawbacks of the techniques will be critically highlighted, showing illustrative case studies generated by the research of Coriolis Pharma together with our scientific board of expertise (Wolfgang Friess, Gerhard Winter, Wim Jiskoot, John Carpenter). One focus of the Archimedes evaluation will be to distinguish protein aggregates from silicone oil droplets with examples from commercial products in pre-filled syringes and model proteins.

3:35 Characterization of Chemical Degradation Products of Peptide Therapeutics by Mass Spectrometry

Peter Nielsen, Ph.D., Scientific Director, Protein Chemistry, Novo Nordisk

Chemical stability is of critical importance in the design and formulation of new peptide therapeutics. We have identified various chemical degradation products in peptide drug candidates. The reactions included deamidation, adduct formation, hydrolysis, and isomerization as well as covalent aggregation. The challenges and strategy for characterization of these modifications using top-down mass spectrometry and ion mobility mass spectrometry will be discussed.

 

Implications of Glass Degradation  

» FEATURED PRESENTATION:  

4:05 Implications of Glass Degradation

Deirdre PiedmonteDeirdre Piedmonte, Ph.D., Principal Scientist in Drug Product Development, Process and Product Development (P&PD), Amgen

This presentation will focus on glass degradation as it affects formulation development. Routes of glass degradation, excipient/buffer and storage conditions that exasperate glass dissolution as well as high throughput analytics useful for formulation screens will be discussed.

4:35 End of Conference



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