Cambridge Healthtech Institute’s 4th Annual
Early Analytical Development for Biotherapeutics
Optimizing the Selection and Performance of Preclinical Analytical Studies
Part of CHI's 8th Annual The Bioprocessing Summit

August 17-18, 2016 | Westin Boston Waterfront | Boston, Massachusetts

The analytical steps conducted in preclinical development following the handoff of a lead candidate are vital on many levels in determining the fate of that program. This complex effort shapes the optimization of the new product, requires the use of expensive and scarce resources and supports the voluminous regulatory filing that is the early IND application. It is imperative that companies reach this important milestone as quickly and efficiently as possible, while positioning the organization to move rapidly into the GMP production needed for early phase clinical trials. Early Analytical Development for Biotherapeutics will present best practice case studies of how industry companies have approached the most important analytical studies occurring during this stage, focusing on the development and optimization of key assays, the application of automation and the challenges of analytical development for novel modalities.

Final Agenda

Wednesday, August 17

7:00 am Registration Opens and Morning Coffee

8:05 Chairperson’s Opening Remarks

Jason Huang, Ph.D., Senior Research Investigator, Bristol-Myers Squibb

Integrated Microfluidic Capillary Electrophoresis-Electrospray Devices for Analysis of Intact mAbs and ADCs

Michael_RamseyJ. Michael Ramsey, Ph.D., Minnie N. Goldby Distinguished Professor of Chemistry, University of North Carolina

Microfluidic devices that include monolithically integrated nano-electrospray emitters are used to electrophoretically separate charge variants of intact monoclonal antibody (mAbs) biotherapeutics and antibody drug conjugates (ADCs). Specialized channel wall coatings allow the use of electrospray friendly buffer systems while maintaining high performance electrophoretic separative performance. Analyses are typically performed in three minutes with little sample preparation while utilizing mass spectrometers with modest resolving power.


9:00 Characterization of Therapeutic Protein by Two Dimensional-LC (2D-LC) System

Shenjiang Yu, Ph.D., Associate Principal Scientist, Analytical Method Development, Merck

Two-dimensional liquid chromatography (2D-LC) is a powerful tool for analyzing highly complex samples such as therapeutic proteins. Both dimensions have orthogonal separation selectivity, thereby increasing the potential peak capacity to the product of the individual peak capacities. With this advanced technology, 2D-LC provides the possibility to combine two orthogonal methods and discover the correlation of the data. This correlation provides critical information for total characterization of therapeutic proteins. .

9:30 Best Practices for Early Stage Bioassay Development

Kari_EfferenKari Sweeney Efferen, Ph.D, Principal Scientist, Pfizer

Incorporation of bioassays is crucial in bioprocess and formulation development since it is often difficult to assess the relevant structure or critical epitopes linked to a particular biological activity using traditional biochemical or biophysical techniques during early development.   It is necessary to ensure that the assay answers the question being asked, such as stability of a critical epitope or relevant structure that can be linked to particular biological activity of that material.

10:00 Coffee Break in the Exhibit Hall with Poster Viewing

10:45 Intact Mass Analysis of Monoclonal Antibodies by Capillary Electrophoresis (CE) – Mass Spectrometry (MS)

Mei_HanMei Han, Scientist, Pharmacokinetics & Drug Metabolism, Amgen, Inc.

Intact mass protein characterization provides important sequence integrity and post-translational modification information. While LC-MS is widely used in the biopharmaceutical industry, CE-MS is an attractive alternative for its high separation efficiency, sensitivity, and minimal sample carryover. However, the intrinsic technical difficulty has hindered the application of this technique. Here, we report the implementation of a recently developed CE-MS interface for proteins from predose and postdose pharmacokinetic samples characterization..

11:15 Ion-Pairing Effect on the Behavior of Synthetic Peptides in Reversed-Phase Liquid Chromatography

Jason_HuangJason Z. Huang, Ph.D., Senior Research Investigator, Bristol-Myers Squibb

We have conducted a systematic study of ion pairing reagents for the reversed-phase HPLC analysis of therapeutic peptides with the goal of improving impurity separations. We studied the influence of ion pairing reagents TFA, NaClO4 and KPF6 on retention and peak shape of peptides on RPLC. As a model system several cyclic and linear peptides with a wide range of molecular weight, polarity and charge were studied..

11:45 Increasing Analytical Throughput for Late Phase Process Development - An Automation Approach for Sample Management

William Grimm, Associate Scientist, Process Development & Analytics, Bristol-Myers Squibb 

A liquid handling system has been configured to act as a sample transfer and dilution platform that can dynamically organize samples on 96-well plates. Automation of ELISA assay setup is handled by a combination of software scripts and Excel VBA commands, this allows for a simple spreadsheet to be used by the analyst to setup a run. Routine use of this technology for sample preparation and organization is invaluable for projects in an accelerated drug development timeline.

12:15 Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own

1:00 Session Break


1:45 Chairperson’s Remarks

Udayanath Aich, Principal Scientist, Process Analytics, Biopharmaceutical Development, Sanofi-Genzyme

1:50 Application of DoE to Optimize Product and Process Parameters

Steven_LaBrenzSteve LaBrenz, Ph.D., Scientific Director, Janssen R&D LL.

The adoption of Quality by Design during drug product development as a regulatory requirement may necessitate the use of DoE in the laboratory to satisfy development activities. Multivariate analysis can be accomplished in many different experimental designs (Taguchi, traditional, definitive), as well as modeling approaches (PLS, PCA). To gain the full benefit of a DoE, consideration needs to be given to the normalized form of data from which means and limits can be derived.

2:20 Recent Advances in Process Analytics for High-throughput Analysis of N-Glycans from Therapeutic Recombinant Proteins

Udayanath_AichUdayanath Aich, Principal Scientist, Process Analytics, Biopharmaceutical Development, Sanofi-Genzyme

With the increase of regulatory expectations, the evolution of PAT and emerging manufacturing processes create a demand for robust, sensitive, accurate profiling of protein glycosylation. Potential shortcomings in protein glycosylation profiling include the high hands-on time required for sample preparation and several hours for data acquisition and analysis. This presentation will focus on the state-of-art process analytics for the high throughput sample preparation and analysis of N-glyans from therapeutic proteins.

2:50 High Throughput Assays for Evaluation of Post-Translational Modifications

Andras_GuttmanAndras Guttman, Ph.D., Senior Manager, SCIEX; MTA-PE Lendulet Professor of Translational Glycomics, Horváth Csaba Laboratory of Bioseparation Sciences, University of Debrecen, Hungary

With the sharp increase in the number of approved protein therapeutics, comprehensive characterization of these new generation drugs is crucial for the biopharmaceutical industry and regulatory agencies. Biotherapeutics possess various post-translational modifications (PTMs) and prone to potential degradation hotspots during the manufacturing process that may affect their efficacy and immunogenicity. This presentation will cover the state of the art of high throughput separation methods for structural elucidation of protein modifications.

3:20 Application of HT Electrophoresis in Early Biosimilar Development

Urška_KristanUrška Kristan, Ph.D., Researcher and Analyst, Analytical Development, Sandoz Biopharmaceuticals, Slovenia

Current initiatives in biosimilar development require thorough understanding of the relationship between product quality and process parameters. In order to achieve this it is necessary to analyze many samples in a short time, therefore HT analytics is often applied in early development. Microchip capillary electrophoresis is one of the HT techniques with many advantages but also some disadvantages, which will be presented in this talk.

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

Thursday, August 18

8:00 am Registration Opens and Morning Coffee


8:25 Chairperson’s Remarks

Joel Bard, Ph.D., Principal Research Scientist, Pfizer

8:30 Analytics by Design: Early Stage Method Development Guided by Molecular Features

Felix_SchumacherFelix Schumacher, Ph.D., Senior Scientist, Large Molecule Research, Roche Diagnostics GmbH, Germany

“Analytics by Design” is a novel concept for early stage analytical method development. Here, analytical packages are tailored to the critical quality attributes of next-generation biotherapeutics, in contrast to the use of generic platform methods. A streamlined method development is based on theoretical considerations and data collected during developability assessment. Such an approach yields valuable insights into multiple aspects of early stage process development, which may guide CMC considerations of future project stages.

9:00 Mass Spectrometric Analysis of Novel Antibody-Drug Conjugates (ADCs) in a Fast-Paced Environment

Lintao_WangLintao Wang, Ph.D., Principal Scientist and Mass Spectrometry Group Lead, Analytical and Pharmaceutical Science, ImmunoGen, Inc.

Designing a safe and effective ADC involves extensive screening of various antibody, linker and payload combinations, which require efficient analysis of a large number of samples with different physicochemical properties. This presentation will focus on the capabilities of size-exclusion chromatography coupled with electrospray mass spectrometry (SEC-MS), a platform assay that is used for generating important information about quality attributes of both antibodies and ADCs in a fast-paced developmental environment.

9:30 A Combination of Structural and Empirical Analyses Delineates the Key Contacts Mediating Stability and Affinity Increases in an Optimized Biotherapeutic Single-Chain Fv

Joel_BardJoel Bard, Ph.D., Principal Research Scientist, Pfizer

Single-chain Fv proteins are key building blocks of bispecific therapeutic antibodies. The causes of scFv instability problems remain poorly understood. We performed detailed structural and biochemical analyses to understand the mechanisms that confer both tighter binding and improved stability on the result of a phage display optimization campaign. This work demonstrates that, aside from being the critical mediators of target binding, CDRs may also be primary drivers of biotherapeutic developability.

10:00 Coffee Break in the Exhibit Hall with Poster Viewing

10:45 Characterization of Formulation Excipient Using Thermal and Spectroscopic Methods for Robust Drug Product Development

Rushikesh_PatelRushikesh Patel, Associate Research Scientist, Bristol-Myers Squibb

Excipient performance during processing can have a significant impact on critical quality attributes of the product. Therefore, detailed characterization of excipient behavior is critical during early development and process design. Here, we present application of multiple thermal and spectroscopic methods for understanding phase behavior of trehalose, a commonly used stabilizer, during freeze/thaw process. The impact of phase change on product quality of a fusion protein is also highlighted.

11:15 A Novel Approach to Isolate and Characterize Product Variants in a Monoclonal Antibody

Xiaoxiao_LiXiaoxiao Li, Ph.D., Senior Scientist, Analytical Development, Abbvie Biotherapeutics

Characterization of binding potency of product variants is required as part of the well-characterized biotherapeutic paradigm. Here we present a novel “divide and conquer” approach to directly isolate variants that differ in binding potency. This approach is applicable to a wide range of biotherapeutics, requires no prior knowledge of the variants, and enables rapid identification and characterization of all variants that differ in binding potency from the main product.

11:45 A Chromatographic Method for Quantifying Individual Monoclonal Antibodies (mAb) in a Co-Formulated Solution

Lin_LuoLin Luo, Research Associate, Formulation Development, Regeneron Pharmaceuticals, Inc.

Developing a method to separate and quantify individual mAb’s from a mixture of multiple monoclonal antibodies is challenging because the mAb molecules may have similar molecular weights, protein structures and molecular charges. In this case study, we present our approaches to developing a quantification method for a co-formulation of three different monoclonal antibodies. In addition, the advantages and limitations of the different methods will be discussed.

12:15 Lunch Available for Purchase in the Exhibit Hall

1:15 Dessert Refreshment Break in the Exhibit Hall with Poster Viewing

1:55 End of Conference