Cambridge Healthtech Institute’s 4th Annual

Advances in Characterization Methods & Approaches

Harnessing Analytics to Speed Development

March 5-6, 2019 * The Westin Alexandria * Alexandria, VA

 

Today’s new frontier of targeted therapeutics includes engineered proteins and live modalities, from antibody-based constructs such as bi or multi-specifics, ADCs to fusion proteins, cell and gene therapies.  How we advance this multi-modality pipeline will depend on the product quality attributes of the molecules.

The 4th Annual Advances in Characterization Methods & Approaches conference provides perspectives and case studies on analysis of structure-function, PTM, conformation and product quality attributes, methods to integrate MAM and data science into analytics, as well as analytical strategies for process control and monitoring.

Final Agenda

 

TUESDAY, MARCH 5, 2019

1:00 pm Registration

1:40 Chairperson’s Opening Remarks

Michael Butler, PhD, CSO, Cell Technology, National Institute of Bioprocessing Research & Training (NIBRT)

CHARACTERIZING NOVEL & COMPLEX BIOTHERAPEUTICS

1:50 FEATURED PRESENTATION: Regulatory Considerations in Developing Analytical Strategies for Gene Therapy Products

Zenobia F. Taraporewala, PhD, CMC (Product) Reviewer, Gene Therapies Branch, Division of Cellular and Gene Therapies, Office of Tissues and Advanced Therapies, CBER, FDA

 

 

2:20 Understanding Structure and Function of Adeno–Associated Virus Based Gene Therapy

Jin_XiaoyingXiaoying Jin, PhD, Senior Principal Scientist, Sanofi

Viral capsid proteins play an important role in cellular targeting and trafficking as part of the viral infection cycle, and thus any changes in the viral capsid protein sequence or post-translational modifications (PTMs) might impact viral targeting and infectivity. We evaluated the role of AAV capsid protein PTMs on AAV transduction potential by generating AAV2 and AAV5 capsid mutants and stress study.

2:50 A Generic mAb Subunit LC-MS Assay for in vivo Drug-to-Antibody Ratio, ADC Quality Attribute Monitoring, & Concentration Determination in Pre-Clinical Studies

Kellie_JohnJohn F. Kellie, PhD, Associate GSK Fellow and Investigator, Bioanalysis, Immunogenicity, and Biomarkers, GlaxoSmithKline

We present the development and details of a single, generic, anti-human mAb LC-MS approach to determine in vivo Drug-to-Antibody Ratios (DARs), pharmacokinetic (PK) concentration, and biotransformation/quality attribute monitoring. Here, information is gained in a single assay for downstream decision making or follow-up testing, and the assay can be utilized for any ADC with a human mAb in any pre-clinical species.

3:20 Automating Data Analysis for the Characterization of Biotherapeutics Candidates

Tomazela_DanielaDaniela Tomazela, PhD, Associate Principal Scientist, Biology-Discovery, Mass Spectrometry, Merck

As part of discovery-developability, sequence-variant evaluation and analysis of stressed samples are critical steps on the path toward final candidate selection. The diverse and large volumes of data generated during the evaluation of molecules’ intactness, sequence coverage, and potential liabilities makes producing reports cumbersome and time-consuming. In this talk, we will focus on MS analysis of biotherapeutic candidates to demonstrate an implementation of data analysis automation in support of early-discovery.

3:50 Refreshment Break in the Exhibit Hall with Poster Viewing

4:30 Bioanalytical Techniques for Bispecific Antibodies and Fusion Proteins

Cook_KevinKevin Cook, PhD, Senior Scientist, Pharmacokinetics & Drug Metabolism, Amgen, Inc.

As biologic therapeutics become more complex, the analytical approaches used to track their pharmacokinetics must also evolve. When targeting multiple pathways, understanding the stability of each targeting arm is crucial to interpret experimental outcomes. This talk will highlight the methods we have used to characterize bispecific antibodies and fusion proteins, while also introducing new methods that can predict stability and reduce the in vivo study burden.

5:00 Cytof/Mass Cytometry Analytics for Cellular Drug Products – Opportunities and Challenges

Katja Kleinsteuber, PhD, Scientist II, Cellular Analytics, bluebird bio

Cell-based gene therapy drug products are a heterogeneous mixture of phenotypically and functionally distinct cells. Complete phenotypic composition analysis requires tools that efficiently deconvolute cellular subpopulations of these drug products. To that aim, we employ high-dimensional mass cytometry (CyTOF) to reliably quantify the phenotypic composition of autologous CD34 and CAR-T drug products. In this talk we present the application of this cutting-edge technology to better understand the composition of our drug products and summarize strategies to overcome common challenges inherent to this technology. 

5:30 Bioanalytical Characterization of an Engineered E.coli Nissle Therapeutic from Discovery to the Clinic

Castillo_MaryMary Castillo, PhD, Lead Metabolic Analytical Scientist, Bioanalytical Research, Synlogic Inc.

The development of Synthetic BioticTM medicines uses an innovative platform where genetically engineered bacteria are designed to respond to the gastrointestinal environment and perform metabolic functions. One engineered strain, SYNB1618 has been designed as a potential therapeutic to lower toxic phenylalanine levels observed with the metabolic disorder, Phenylketonuria.  Here we demonstrate the methodologies used to assess safety and efficacy of an engineered strain of SYNB1618 for lowering phenylalanine levels and producing biomarkers to show the strain activity, biosafety and pharmacokinetics.

6:00 End of Day/Short Course Registration

6:30 - 9:00 Dinner Short Courses*

 

WEDNESDAY, MARCH 6, 2019

8:00 am Morning Coffee

8:30 Chairperson’s Remarks

Bruce Yu, PhD, Professor, Pharmaceutical Sciences, University of Maryland School of Pharmacy

CHARACTERIZATION OF STRUCTURE-FUNCTION, PTM AND QUALITY ATTRIBUTES

8:35 High-Resolution Mass Spectrometry Characterization of Post-Translational Modifications (PTMs) and Sequence Extensions in Fc-Fusion Proteins

Spahr_ChrisChris Spahr, Senior Scientist, Discovery Attribute Sciences, Amgen, Inc.

High-resolution mass spectrometry (HR-MS) is an invaluable tool for characterizing PTMs and sequence extensions. O-xylosylation has been observed in the (G4S)n sequences of protein linkers in Fc-fusion proteins. Hydroxyproline (Hyp) has been identified in linker sequences resembling collagen, while 3-Hyp and 4-Hyp isomers may be differentiated using ETD/HCD (electron-transfer dissociation/ higher-energy collision dissociation). Finally, de novo sequencing, enabled by HR-MS/MS has elucidated sequence extensions of Fc due to aberrant splicing. Examples will be provided.

9:05 Intact and Subunit Level Approaches for Enhanced CQA Assessment and Structure-Function Characterization of Biotherapeutics

Graf_ChristianChristian Graf, PhD, Principal Scientist, Technical R&D, Technical Dev Biosimilars, Novartis Pharma AG

Here, we will present advanced analytical methods to characterize and quantify critical quality attributes like PTMs of therapeutic antibody candidates. Novel chromatographic approaches, such as HILIC and polyphenol RP-UPLC, coupled to high-resolution mass spectrometry enabled detailed PTM characterization on intact mAb and subunit level. Additionally, intact or subunit HDX-MS methods provide rapid and robust information about the impact of modifications on the global higher order structure. Case studies on clinical antibody candidates will be shown.

9:35 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.

Challenges with Characterizations of Antibody-Drug Conjugates

Moderator: Zhiqi Hao, PhD, Scientist, Protein Analytical Chemistry, Genentech

  • Current analytical technologies used to perform ADC Characterization
  • Technical challenges due to the complexity associated with ADCs and how the challenges are addressed
  • Application of LC-MS (MS/MS) on ADC characterization
  • Phase of ADC development and regulatory requirement of characterization

Techniques for Characterizing Novel Molecules – Bispecifics and Fusion Proteins

Moderator: Kevin Cook, PhD, Senior Scientist, Pharmacokinetics & Drug Metabolism, Amgen, Inc.

  • How many assays are needed when a therapeutic has multiple targets?
  • What are the benefits/weaknesses of “intact” and “total” assays?
  • How to choose assay platform between LBA and LC-MS?

Strategies and Considerations for Characterization of Gene Therapy Products

Moderator: Xiaoying Jin, PhD, Senior Principal Scientist, Sanofi

  • Characterization strategy due to material constraint
  • General biochemical/biophysics characterization for gene therapy characterization
  • LC-MS characterization/application

 

10:35 Coffee Break in the Exhibit Hall with Poster Viewing

11:15 Structural Characterization of Antibody-Drug Conjugates

Hao_ZhiqiZhiqi Hao, PhD, Scientist, Protein Analytical Chemistry, Genentech

During clinical development of ADCs, it is critical to characterize product structure-function, and understand the impact of product quality attributes and variants on safety and efficacy. This presentation focuses on using comprehensive and novel analytical techniques to characterize ADCs produced with different linker-drug and conjugation chemistries. Case studies include analyzing conjugated peptides, rare product variant, and drug degradation pathways. Novel mass spectrometry methods that provide automatic and comprehensive characterization will be highlighted.

11:45 Microchip Electrophoresis Coupled to MS for Rapid Product Quality Monitoring

Madren_SethSeth Madren, PhD, Scientist I, Analytical Development, Biogen

Therapeutic mAbs are highly heterogeneous, and multiple product quality attributes of mAbs need to be monitored or characterized to assure product safety and efficacy. Here, we present a HT CE-MS method that analyzes IdeS digested mAb in cell culture medium or drug substance, that enables near-real time monitoring of multiple product quality attributes of mAb product at domain level. This method also presents unique advantage of monitoring attributes, such as deamidation, that is typically difficult to detect at domain level by traditional LC-MS methods.

12:15 pm Luncheon Presentation: Future Proofing and Understanding Your Molecule with Contemporary Analytics and Formulation Approaches

Greg Adams, PhD, Senior Director, Global Analytical Strategy & Development, FUJIFILM Diosynth Biotechnologies

Topics to be covered include: 1) How contemporary analytical tools help us understand the personality of a protein 2) Approaches to balancing science and the need to rapidly get new molecules into the clinic and 3) Tools we use to ensure we are employing to "future-proof" methods for late-phase and commercial use.

12:45 Session Break

1:25 Chairperson’s Remarks

Christian Graf, PhD, Principal Scientist, Technical R&D, Technical Dev Biosimilars, Novartis Pharma AG

NOVEL TECHNOLOGIES AND APPROACHES IN ANALYTICAL STRATEGIES

1:30 FEATURED PRESENTATION: Water Proton NMR for Noninvasive Analysis of Biologics and Other Complex Drugs

Bruce Yu, PhD, Professor, Pharmaceutical Sciences, University of Maryland School of Pharmacy

Water is the dominant component of most pharmaceutical solutions. It is possible to use water proton NMR (wNMR) for nondestructive quantitative analysis of pharmaceutical solutions. Examples of using wNMR to monitor protein aggregation, amide hydrolysis and nanoparticle clustering will be presented. Data will also be presented on FDA-approved drugs, both biologic and non-biologic. Ongoing work focuses developing flow-wNMR as an in-line process analytic technology for continuous biomanufacturing.

2:00 Strategic Integration of the Multi-Attribute Method into Early Phase Biotherapeutic Development

Powers_ThomasThomas Powers, PhD, Senior Scientist, Analytical R&D, Pfizer

Pfizer is evaluating the implementation of multi-attribute monitoring (MAM) into early phase biotherapeutic development. Initial efforts have focused on assessing method robustness, evaluating system performance, harmonizing hardware and software, and exploring the capabilities of the resulting data. Importantly, the correlation of MAM results with current release tests has demonstrated the utility of a MAM method for generating consistent results, all while improving sampling throughput. Further implementation of MAM into late phase programs is being evaluated.

2:30 Data Cleaning: Strategies to Generate High Quality Data Sets for Liquid Formulation in a High Throughput Setting

Kathrin Schaeker-Theobald, PhD, Senior Scientist, NBE Formulation Sciences, Abbvie Deutcchland GmbH & Co. KG

Regulatory guidance requests multivariate analysis to define formulation design space and thus to assure safety and efficacy of the product. This requires large and consistent data sets. We will present our data handling approach in our biologics formulation development studies to efficiently filter and clean diverse analytical screening raw data to obtain information-rich data sets usable for in-depth statistical evaluation or data science. Data integrity is a key element that is as well included.

3:00 Advances in LC-MS Based Approaches for Size and Charge Heterogeneity Characterization of Therapeutic mAbs

Shunhai Wang, PhD, Staff Scientist, Analytical Chemistry, Regeneron

Monoclonal antibody (mAb)-based therapeutics are inherently heterogeneous with respect to size and charge due to their complex four-chain structure and the propensity to accommodate multiple enzymatic and chemical post-translational modifications. We present several advances in LC-MS based approaches for size and charge heterogeneity characterization of therapeutic mAbs, including coupling high performance SEC and IEX separation with ultrasensitive native Nano-ESI mass spectrometry detection and a novel HILIC-MS method for comprehensive low molecular weight species profiling.

3:30 Refreshment Break in the Exhibit Hall with Poster Viewing

ANALYTICS FOR PROCESS MONITORING AND CONTROL

4:10 Controlling Glycosylation of Antibodies during Processing

Butler_MichaelMichael Butler, PhD, CSO, Cell Technology, National Institute of Bioprocessing Research & Training (NIBRT)

The enzymic activity profile activity of host mammalian cells as well as their growth conditions determine the glycosylation pattern that result in secreted glycoproteins such as Mabs. The glycosylation profile can be controlled during upstream processing by cell engineering or control of media composition during cell growth. An alternative approach is by enzymic remodeling following harvest from the bioreactor. The relative value of each of these possible strategies will be evaluated for the production of an antibody.

4:40 Translating Advanced Process Control and Process Analytical Technologies into an End to End Control Strategy for Manufacture and Release of Biotechnology Products

Stacey Traviglia, PhD, Associate Director, QC Analytical Technology, Biogen

Recent regulatory draft guidance suggests that a combination of critical process parameters together with an understanding of the product’s critical quality attributes (CQA) may provide greater assurance of product quality than end product testing alone and that product release can be based on a combination of real time release (RTR) and end-product testing. Case studies in which this concept has been applied will be presented.

5:10 Critical Material Attributes and PAT for In-Process Measurement and Control of Product Stability and Consistency

Marina Kirkitadze, PhD, MBA, Head, Process Support and PAT, Sanofi Pasteur Canada

This talk is focusing on the application of inline techniques to measure particle size in real time. The particle sizing measurements were collected on aluminum adjuvants by the focused-beam reflectance measurement (FBRM), particle vision and measurement (PVM) and laser diffraction. The results and next steps are discussed.

5:40 Networking Reception in the Exhibit Hall with Poster Viewing

6:40 End of Day