Archived Content

ANALYTICAL STREAM  

5th Annual  

Protein Aggregation and Stability in Biopharmaceutical Products May 2 - 3


The study and prevention of protein aggregation and its consequences represents one of the most demanding tasks in biomedical research and pharmaceutical manufacturing today. Whether one’s interests lie in protein characterization or analytics, the study of protein aggregation-induced adverse effects or immunogenicity, or work in formulation and process development for protein-based therapeutics, “Protein Aggregation and Stability in Biopharmaceutical Products” will provide a comprehensive real-world perspective on this challenging arena.

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TUESDAY, MAY 1


6:00-9:00 pm Recommended Dinner Short Course* 


*Separate registration required.


WEDNESDAY, MAY 2

7:00 am Registration and Morning Coffee

 

ANALTYICAL METHODS: DETECTION AND CHARACTERIZATION 

8:30 Chairperson’s Opening Remarks

Vineet Kumar, Ph.D., Senior Research Scientist, Global Formulation Sciences, Parenterals, Abbott


» 8:40 Keynote Presentation 

Causes and Control of Aggregation and High Viscosity in High Concentration Formulations

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

At high concentrations, the solution properties of protein formulations are dominated by the colloidal properties of the protein. These colloidal properties arise from weak forces between all of the solution species. The weak forces are electrostatic and electrodynamic in origin, with charge-charge interactions being the longest range and Van der Waals interactions the shortest range force. These weak forces give rise to solution nonideality, a readily measured quantity. Likewise, protein charge is a readily measured quantity that is important to the solution nonideality. This talk will cover: 1) Weak forces and concentrated solutions: definitions, nonideality and nonideality determination; 2) Measuring the charge on proteins: methods, accuracy and meaning; 3) Monoclonal antibody charge: an example of Nature’s perverse sense of humor; 4) Charge and solution behavior: Charge may help or it may hurt solution properties; 5) The role of charge in aggregation; 6) The importance of protein dipole moment measurement.


Sponsored by
Malvern small
9:10 Combining Raman Spectroscopy and DLS for the Physico-Chemical Characterization of Protein Therapeutics

Neil LewisE. Neil Lewis, Ph.D., Chief Technology Officer, Malvern Instruments (biography)

The emergence of protein therapeutics has created a demand for new analytical methods because unlike small molecule pharmaceuticals their efficacy is also determined by their high order structure and conformation. Raman spectroscopy in combination with DLS can be used to measure these properties using small amounts of material under typical formulation conditions.
 

9:40 Characterization of Protein Aggregation in a Fusion Protein

James StrandJames Strand, Staff Research Associate, Analytical Develpment, Acceleron Pharma (biography)

The mechanisms behind non-native aggregation of recombinant Fc-fusion protein therapeutics from mammalian expression systems was studied specifically in relation to highly glycosylated proteins. Both chromatographic as well as higher order structural tools were used to define protein changes that may influence aggregation kinetics. In addition, the impact of glycosylation state on aggregation was also investigated.
 

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

11:10 Sub-Visible Particle Detection in a High Concentration Protein Formulation

Mary Beth PelletierMary Beth Pelletier, Ph.D., Scientist, Analytical Technology, Biogen Idec (biography)

The development and qualification of a method for subvisible particle counting is described for a high concentration protein formulation. Challenges for this method include the concentration of the product and the prefilled syringe container-closure system, leading to the presence of silicon oil droplets in the samples. Significant development was carried out to ensure that the resulting method was robust, reproducible, and suitable for use in a QC environment.

11:40 Novel Mass Spec-Based Approaches to Characterize Aggregation and Association of Protein Therapeutics

Igor KaltashovIgor Kaltashov, Ph.D., Professor, Department of Chemistry, University of Massachusetts, Amherst (biography)

Traditionally, aggregation has been monitored using relatively low-resolution techniques that do not characterize the process at desired detail. An alternative is now provided by electrospray ionization mass spectrometry (ESI MS). We will discuss several recent developments that allow ESI MS to be used both in combination with classical tools and as a stand-alone technique for characterization of both soluble protein aggregates and products of ordered protein association.

Sponsored by
Quanta BioDesign
12:10 pm Luncheon Presentation I
Introducing dPEG®s as a New Paradigm in Addressing Protein Aggregation Issues

Paul D. Davis, Ph.D., President & CEO, Quanta BioDesign, Ltd.

Quanta has generated an abundance of data in working with proteins which suggests that its dPEG® technology could be a simple, yet flexible solution to solving or affecting many of the aggregation issues that are extant in biologics. In support, there are a number of key published references using the dPEG® technology to solve a wide range and variety of scenarios related to solubility, aggregation and elimination of non-specific binding.  Data demonstrating these positive attributes of dPEG®s, as well as data on binding/activity and immunogenicity will be discussed.

12:40 Luncheon Presentation (Sponsorship Opportunity Available)

 

Understanding Mechanisms Of Aggregation: Extrinsic Factors And Excipients 

1:30 Chairperson’s Remarks

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




 

1:35 Mechanistic Links between Soluble Aggregate and Subvisible Particle Formation in Protein Therapeutics

Robert Simler, Ph.D., Staff Scientist, BioFormulations Development, Genzyme (biography)

Although a significant amount of recent work has focused on the quantification and characterization of subvisible particles, little is understood about the mechanism which links their formation to that of smaller, soluble aggregates. This talk will focus on agitation experiments designed to generate subvisible particles and soluble aggregates in an effort to identify a mechanistic relationship between the two higher order species. Practical consequences of this mechanism as they relate to formulation development will also be discussed.


» 2:05 Featured Presentation 

High Concentration Protein Formulations: Complexity of the Stabilization and/or Destabilization Effects of Sugars/Polyols on Aggregation

Vineet KumarVineet Kumar, Ph.D., Senior Research Scientist, Global Formulation Sciences, Parenterals, Abbott (biography)

Sugars are widely used in low/high concentration protein formulations. In fact, more than 70% marketed protein formulations have sugars as stabilizers and/or tonicity modifiers. The talk will focus on the stabilization/destabilization effects of sugars and the strategies to select the best and the most useful poyol for the formulation. Audience will learn methods to appropriately select and/or deselect sugars especially for high concentration formulations.


2:35 Recent Advances in Protein Size and Formulation Viscosity Measurements

Kevin Mattison, Ph.D., Principal Scientist - Bioanalytics, Malvern Instruments (biography) - Co-authored by: Hanna Jankevics, Product Technical Specialist, Malvern Instruments and David Goodall, CSO, Paraytec Limited

Two formulation properties important to achieving the Commercial Target Profile for biotherapeutics are protein size and formulation viscosity.  For concentrated mAb formulations, reduction of the viscosity to a level facilitating subcutaneous injection is critical, and often very challenging, due in large part to the minimum volume requirements associated with rheological testing.  The objective of this presentation is to describe recent technological innovations providing automated measurements of protein size and formulation viscosity with sample volumes of less than 7 microliters.

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


3:50 Problem Solving Breakout Discussions

Concurrent problem solving breakout discussions, open to all attendees, speakers, sponsors, and exhibitors, provide a forum for discussing key issues and meeting potential collaborators. Plan to take part and explore these topics in-depth. Please pick a topic of your choice, find your table and join in.

Opalescence Phenomenon of Protein Therapeutics Formulated at High Concentrations

Moderator:  Yin Luo, Ph.D., Director, Mass Spectrometry and Biophysical Characterization, Analytical Research & Development, BioTherapeutics Pharmaceutical Sciences, Pfizer, Inc.

• How common is the opalescence phenomenon among high-concentration protein therapeutics?
• Are there commonalities in protein attributes among molecules that are opalescent?
• Is protein opalescence caused by protein aggregation?
• Does protein opalescence share similar molecular mechanisms with protein aggregation?
• How to address potential immunogenicity concerns for the opalescent protein therapeutics?
• What characterizations are needed to understand the phenomenon of protein opalescence?
• Are the necessary technologies to understand protein opalescence available yet?

The Effect of Sugars on Protein Stability

Moderator:  Devendra (Davy) Kalonia, Ph.D., Professor of Pharmaceutics, Department of Pharmaceutical Sciences, University of Connecticut

• Mechanism of sugar stabilization of proteins
• Effect of sugar molecules on the propensity of protein molecules to stay in solutions: solubility
• Opposite effect of sugars: enhanced destabilization and aggregation of proteins because of mechanical stress

Assessing 'High Order Structure' of Protein Therapeutics

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

• What role do changes/modifications of higher order (secondary, tertiary) protein structure play in aggregation?
• Can monitoring relative changes in higher order protein structure in-vitro using optical spectroscopy (CD, FTIR, Raman, etc.) provide rapid insight into optimal formulation conditions and predict long-term product stability?
• Can monitoring relative differences in protein higher order structure in-vitro using optical spectroscopy (CD, FTIR, Raman) provide useful comparative data for assessing biosimilarity?

Causes and Control of Aggregation and High Viscosity in High Concentration Formulations

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

• Weak forces and concentrated solutions: definitions, nonideality and nonideality determination
• Measuring the charge on proteins: methods, accuracy and meaning
• Monoclonal antibody charge: an example of Nature’s perverse sense of humor
•  Charge and solution behavior: Charge may help or it may hurt solution properties
• The role of charge in aggregation
• The importance of protein dipole moment measurement

Sub-Visible Particle Analysis and Characterization During Product Development

Moderator:  Dingjiang (Dean) Liu , Ph.D., Fellow Scientist, Formulation Development Regeneron Pharmaceuticals, Inc.

• Accepted and emerging methods
• Comparability challenges during product development
• Mechanism of particle formation

Aggregate in Formulations and Their Potential Risk in Induction of Immunogenicity

Vibha Jawa, Ph.D., Principal Scientist, Clinical immunology, Amgen, Inc.

• Biophysical attributes and their role in induction of immune response
• Innate, adaptive and bystander cell activation
• Activation thresholds and their influence (type of molecule, donor status, mode of administration, etc.) for induction of immune response
 

4:50-6:00 Networking Reception in the Exhibit Hall with Poster Viewing
Emerald BioStructuresPoster Awards Sponsored by



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