Cambridge Healthtech Institute’s 3^rd Annual

Method Development, Qualification & Validation

Developing Fit-for-Purpose Methods throughout the Product Lifecycle

March 4-5, 2019 * The Westin Alexandria * Alexandria, VA

From method development and qualification to validation and transfer, it is a continuous yet interdependent process from early R&D to QC and release; the objective is to prove that methods selected are fit for their intended purpose. With an expanded portfolio of newer formats and non-mAbs in the pipelines, knowledge, experience and advanced techniques are critical to establish efficient, accurate and fit-for-purpose analytical methods that can be subsequently qualified, validated and transferred to release testing, thereby improving efficiency and reducing cost, time and resources.

The 3rd Annual Method Development, Qualification & Validation conference highlights methods for complex molecules, methods to determine conformational and stability of therapeutic proteins, statistical approaches to validating methods, as well as strategies to meet regulatory requirements.

Final Agenda

MONDAY, MARCH 4, 2019

7:30 am Registration and Morning Coffee

8:30 Chairperson’s Opening Remarks

Marla Abodeely, PhD, Head, Bioassay/Immunoassay Method Development, Shire Pharmaceuticals

DEVELOPING METHODS FOR COMPLEX AND EMERGING MOLECULES

8:40 Development of Methods for the Characterization of Adeno-Associated Virus Vectors

Zhongying Chen, PhD, Senior Scientist, Analytical Bioassay Development, MedImmune

Adeno-associated virus (AAV) provides a promising viral vector platform for gene therapy. Analytical characterization of virus products requires comprehensive method development to assess virus identity, potency, purity, safety and stability. This presentation will discuss the development of methods to analyze virus genome copy number, transduction and infectivity titer, potency and biophysical characteristics in support of product development from upstream vector screening to downstream purification and formulation development.

9:10 Genome Content and % Full Capsid Analysis of Adeno-Associated Viruses by Spectrophotometry

Keith Webber, Senior Scientist, Analytical Development, RegenxBio

A spectrophotometry method was developed for the fast and reliable measurement of genome content, capsid content, and empty/full ratio of adeno-associated viruses (AAV). Spectrophotometry is a common technique for measuring concentration of biological products, and the technique was applied to AAV for its two major components (protein and DNA). This method has demonstrated to be precise and accurate with comparable results to other conventional methods, including PCR, TEM, and AUC.

9:40 The Development of a Sensitive and Quantitative Enzyme Activity Biomarker Assay to Support Gene Editing Clinical Trials for the Treatment of a Lysosomal Storage Disease

Liching Cao, Director, Bioanalytical Operations, Sangamo Therapeutics, Inc.

Clinical studies involving gene therapy to treat LSD patients with enzyme deficiency utilize an enzyme activity assay as one of the biomarker assays for efficacy evaluations. The commercially available diagnostic test is a non-quantitative enzymatic activity assay that uses an artificial fluorescent chemical as standard. We developed a quantitative assay by incorporating recombinant enzymes as reference standard and controls, with assay sensitivity capable of quantitating enzyme activity in both diseased and normal ranges.

10:10 Networking Coffee Break

10:40 CGE Purity and Characterization Method Development for Multiple Virus Like Particle (VLPs) Vaccines

Deepika Gollapudi, MSc, Scientist, Analytical Development, National Institutes of Health

A new method was developed as a high-throughput with a highly sensitivity for purity determination and characterization of virus-like particle (VLP) vaccines based on PerkinElmer GX II (CGE). The method was qualified and shown to be accurate, repeatable (less than 2% relative standard deviation of the mean (% RSD) for both intra and intermediate precision). This new method was successfully applied for lot release, as well as for tracking the vaccine lot to lot variations. 

11:10 Developability Assessment for Candidate Selection

Gunasekaran Kannan, PhD, Senior Director, BioTherapeutics, Denali Therapeutics, Inc.

11:40 Developing a Robust icLEF Platform Method

Danielle Hamler, Principal Research Associate, BioProcess Analytics, Sanofi

Development of a robust harmonized platform method across global sites within a company can have significant impact in cost reduction and speed of manufacture. We gathered a global analytics team to develop a harmonized high throughout (HTP) method for cIEF. The team utilized risk assessment tools (Fishbone diagram) and JMP analysis software to ensure the harmonized method was robust. This presentation will focus on the development of the harmonized method and the robustness studies performed to achieve a HTP harmonized cIEF method.

12:10 pm Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

12:40 Session Break

1:40 Chairperson’s Remarks

Holly London, PhD, Scientist II, QC, Biomarin Pharmaceuticals

REGULATORY CONSIDERATIONS AND Method LIFE CYCLE MANAGEMENT

1:45 Regulatory Considerations for Analytical Method Development to Support Expedited Development Programs

Justin Alridge, PhD, RAC, Senior Manager, Global Regulatory Affairs CMC, Biogen

2:15 Method Development and Life Cycle Management in a Fast-Paced Environment

Holly London, PhD, Scientist II, QC, Biomarin Pharmaceuticals

2:45 Novel Analytics to Improve Bioprocessing and Validation Guideline Review

Fredrik Sundberg, Global Director, Strategic Customer Relations, GE Healthcare Life Sciences

A thorough understanding of how the process impacts the product quality attributes is essential for decision-making. Implementation of a robust analytical control strategy with information-rich technologies has the potential to significantly improve characterization and productivity. This presentation will provide an overview of how innovative protein analysis tools have been adopted for better comparability, release testing and impurity monitoring. It will also cover the latest regulatory strategies including the new SPR validation guidelines.

3:15 Networking Refreshment Break

3:45 BREAKOUT DISCUSSION SESSION

Join your peers and colleagues in facilitated, small-group discussions to exchange ideas, share best practices, discuss challenges and make new contacts.

Protein Footprinting - HDX, FPOP and Other Approaches

Moderator: Michael L. Gross, PhD, Professor, Chemistry, Immunology and Medicine, Washington University in St. Louis

• Advantages and disadvantages of HDX
• Advantages and disadvantages of FPOP
• FPOP and other approaches vs. Synchrotron footprinting
• Role of other footprinting approaches (e.g., NEM, GEE, BHD)

Challenges in Establishing Bioassay Acceptance Criteria and Bioassay Method Validation & Transfer

Moderators:

Liming Shi, MS, MA, Executive Director, Quality Control, CMAB Biopharma Inc. and

Marla Abodeely, PhD, Head, Bioassay/Immunoassay Method Development, Shire

• Stage appropriate acceptance criteria set up along with method development
• Accumulation of so-called "enough" historical bioassay data
• Both scientific and statistically sound bioassay acceptance criteria
• Challenges in bioassay method validation and transfer

Analytical Tools and Techniques for Host Cell Protein Characterization

Moderator: Christopher Yu, PhD, Principal Scientist, Protein Analytical Chemistry, Genentech, Inc.

Computational Re-design of Protein Solubility

Moderator: Salvador Ventura, PhD, Director, Institute of Biotechnology and Biomedicine, Autonomous University of Barcelona

• The role of protein sequence
• The role of protein structure
• The role of protein stability
• The role of exrinsic factors

JOINT PLENARY SESSION

4:45 Keynote Introduction

Katelyn Smith, PhD, Senior Scientist, Pharmaceutical Sciences, Merck & Co., Inc.

4:50 KEYNOTE PRESENTATION I: Mass Spectrometry for Higher Order Structure and Properties of Therapeutic Proteins

Michael L. Gross, PhD, Professor, Chemistry, Immunology, and Medicine, Washington University in St. Louis

We are developing methods for protein therapeutics to determine higher order structure and to assure the structure has not changed. Our methods bring understanding to hidden conformations that characterize mutant proteins from bacterial and viral sources and to amyloid-forming proteins. Of additional interest are methods for determining protein affinity with small molecules (drug candidates). The measurement strategies are mass-spectrometry-based protein footprinting (HDX, FPOP, and other labeling strategies) and native MS with ion mobility.

5:25 KEYNOTE PRESENTATION II: Combining Structural Aggregation Propensity and Stability Predictions to Re-Design Protein Solubility

Salvador Ventura, PhD, Director, Institute of Biotechnology and Biomedicine, Autonomous University of Barcelona

The aggregation propensity of each particular protein seems to be shaped by evolution according to its natural abundance in the cell. The production and downstream processing of recombinant polypeptides implies attaining concentrations that are orders of magnitude above their natural levels, often resulting in their aggregation. We demonstrate that our AGGRESCAN 3D structural aggregation predictor allows designing mutations at specific positions in the structure that improve the solubility of proteins without compromising their conformation. Our data indicate that the solubility of unrelated proteins can be easily tuned by in silico-designed non-destabilizing amino acid changes at their surfaces.

6:00 Welcome Reception in the Exhibit Hall with Poster Viewing

7:00 End of Day

TUESDAY, MARCH 5, 2019

8:00 am Morning Coffee

8:30 Chairperson’s Opening Remarks

Danny K Chou, PhD, President, Compassion BioSolution, LLC

METHODS FOR STABILITY ASSESSMENT OF THERAPEUTIC PROTEINS

8:35 Development of Quantitative and Stability Indicating Methods for Polysorbate 80

Ying Xu, PhD, Scientist, Biologics Development, Sanofi

A simple and reproducible HPLC method for quantification and stability of polysorbates has been developed and validated. No or minimal sample preparation is needed for biotherapeutics. The quantification method was capable of reporting polysorbates as low as 10ppm and the sensitivity could be further improved on a needed basis. This method was applied to multiple drug modalities including proteins, monoclonal antibodies and adeno-associated viruses.

9:05 Combining Orthogonal Methods with DOE to Achieve the Most Robust Biopharmaceutical Product Design in Less Time

Danny K Chou, PhD, President, Compassion BioSolution, LLC

The objective of this presentation is to demonstrate some practical ways one can go about measuring both conformational and colloidal stability of a protein and use this information along with real-time aggregation data to choose the most suitable solutions for improving both thermal and interfacial stability of a protein molecule. Part of the emphasis will be on how to combine Experiment by Design (EbD) with innovative analytical technologies to obtain data with efficiency and the statistical power to enable decision making.

9:35 A Tale of Three Transfers: The Good, the Bad, and the Ugly - Transfer of mAb Purity Analysis to cGMP Organizations

Niomi Peckham, Development Scientist III, Alexion Pharmaceuticals Inc.

Alexion’s focus on rare diseases translates into accelerated development timelines, making high throughput and flexible platforms a necessity. Protein analysis on the PerkinElmer LabChip® instrument has been adopted by Alexion for monoclonal antibodies from Pre-Clinical through Validated Phase II/III release and stability applications. Contract manufacturing organizations have also been utilized for manufacturing and analytical support to progress several clinical stage molecules, resulting in repeated transfers of validated assays to cGMP laboratories.

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

METHOD VALIDATION & TRANSFER

10:45 Statistical Approaches for Method Validation

Jianchun Jason Zhang, Senior Principal Statistician, MedImmune

Analytical methods are utilized throughout drug research and development to aid quality decision-making. Although several statistical test methods have been proposed in literature to test the “fit for use” hypothesis, the majority of the methods are not designed to protect the risk of accepting unsuitable methods, thus having the potential to cause uncontrolled consumer’s risk. In this talk, a method based on the concept of total error and generalized confidence interval is presented.

11:15 The Upside of a Difficult Method Transfer

Marla Abodeely, PhD, Head, Bioassay/Immunoassay Method Development, Shire Pharmaceuticals

This presentation will cover the trials and tribulations of transferring a late-stage, cell-based potency method to a quality laboratory for a complex biologic.

11:45 Challenges in Analytical Method Transfer: Global View

Kuruppu Dharmasiri, PhD, Senior Scientist, Global Technical Operations, AstraZeneca Biologics

With the development and commercialization of life-saving biotechnology products, analytical method transfers are becoming an essential component of the current global biotechnology environment. There are differences between internal transfers and external transfers to global sites. In this presentation, practical challenges encountered during method transfers are discussed with examples.

12:15 pm Close of Conference