Advances in Characterization Methods and Approaches

With newer and more complex molecules taking the spotlight these days, companies need better and faster ways to characterize these non-traditional antibodies. These new entities, coming in the different flavors of bispecifics, conjugates, multi-components, combinations, fusions etc., do not work well with the existing platforms. Coupled with the massive amount of data coming out from mass spectrometry and the regulatory demands for more accurate particle analysis and immunogenicity risk assessment, the industry is eager for novel approaches, proven methods and creative strategies that can help them analyze, interpret and translate information into innovation.

The Advances in Characterization Methods and Approaches conference arms scientists with the tools to develop the right strategies to speed up innovation, such as developing high throughput techniques for multi-parameter analysis; using in-process control and real-time release testing to improve CMC process; improved methods for biochemical and biophysical analysis, as well as advanced techniques for better understanding of the molecule’s structure-function dynamics, charge heterogeneity, post-translational modifications, and sequence variants.

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12:30 pm Registration

1:30 Chairperson’s Opening Remarks

Baolin Zhang, Ph.D., Senior Investigator & Product Quality Reviewer, Office of Biotechnology Products, Center for Drug Evaluation and Research, U.S. FDA (invited)



1:40 Real-Time Release and Multi-Parameter/ Multi-Attribute Analysis

Hans-Martin_MuellerHans-Martin Mueller, Ph.D., Director, BioProcess Development, MSD Merck

The modern biotech industry is constantly striving to modernize the analytical control of processes, drug substances and drug product batches. Today, the analytical characterization should be available earlier, at a higher quality and with more information content. The trend in our industry is to investigate practical approaches for on-line data generation, multi-attribute methods and batch releases in real-time. In biologics drug development the prerequisite for running these new techniques are “platform”-analytical methods, which can be applied to broad range of biologics.  


2:10 High-Throughput Mass Spectrometric Approach for Multi-Attribute Analysis

Yan Wang, Ph.D., Scientist, Analytical Development, Biogen

Peptide mapping is a sensitive technique to monitor a variety of post-translational modifications (PTMs) in a single run. We have recently developed a high-throughput peptide mapping approach to quickly compare multiple protein quality attributes during cell line selection. Our new mass spectrometric approach has enabled us to quickly compare product quality attributes among clones. This new workflow is transferrable to assess product quality not only for cell line selection, but also for manufacturing and process development.

2:40 Advancing Analytical Characterization for In-Process and Purified Protein Pharmaceuticals

Anastasiya_ManuilovAnastasiya Manuilov, MS, Senior Scientist, Analytical Research and Development, Pfizer

The development of analytical methods for the characterization of protein biopharmaceuticals faces complex challenges. 2D-LC method development will be presented for the effective separation of IgG1 and IgG2 isoforms, via pH-gradient cation exchange with an in-line protein-A column. This setup allows minimal sample preparation for direct analyses of conditioned cell culture media and for characterization of various product isoforms.

Bruker3:10 Presentation to be Announced

3:40 Refreshment Break in the Exhibit Hall with Poster Viewing

4:10 Monitoring Multiple mAb Attributes Using Peptide Mapping UPLC-MS

Melissa Alvarez, Scientist, Protein Analytical Chemistry, Genentech, Inc.

4:40 High-Throughput Quality Attributes Platform Development

Jasmine Wang, Ph.D., Scientist I, Analytical Biochemistry, MedImmune

5:10 Close of Day Two

5:45 Dinner Short Course*

SC1: SVPs, Aggregates and Impurities: Measurement, Characterization and Impact

SC2: Glycobiology of Antibodies

*Separate registration required.

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8:00 am Morning Coffee

8:30 Chairperson’s Remarks

Hans-Martin Mueller, Ph.D., Director, BioProcess Development, MSD Merck


8:40 Fast Photochemical Oxidation of Proteins (FPOP) Complements HD Exchange for Protein Biophysics and Therapeutics

Michael GrossMichael Gross, Ph.D., Professor of Chemistry, Washington University in St. Louis

We are developing methods for characterization of protein therapeutics to meet the challenge that proteins, unlike small-molecule drugs, can form various higher order structures and can oligomerize. The overarching strategy is mass-spectrometry-based protein footprinting, which has high sensitivity and fast turnaround. We will review HD exchange, specific amino-acid labeling, and fast photochemical oxidation of proteins (FPOP) that meet the challenge to map epitopes, follow oligomerization, and assure that structure of a protein has not changed.

9:10 How Precise Is that Hydrogen-Deuterium Exchange Mass Spectrometry Measurement?

Jeffrey_HudgensJeffrey W. Hudgens, Ph.D., Research Chemist, Biomolecular Measurement Division, Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology

Adoption of hydrogen-deuterium exchange mass spectrometry (HDX-MS) for certifying properties of biotherapeutics cannot occur without a thorough knowledge of its measurement uncertainties. For an international study NIST enlisted experts from the biopharma, non-profit, and academic sectors who conducted HDX-MS measurements on the Fab and Fc domains of the antibody reference material, NIST mAb. Analyses of these data have produced the first determinations of HDX-MS reproducibility for measurements involving mAb materials.


9:40 Potency Assays for Biopharmaceuticals: A Regulatory Perspective

Baolin Zhang, Ph.D., Senior Investigator & Product Quality Reviewer, Office of Biotechnology Products, Center for Drug Evaluation and Research, U.S. FDA (invited)

Because of the complex nature of biopharmaceuticals, it can be scientifically challenging to develop appropriate potency assays for each product. A regulatory evaluation of adequacy of potency assays is made on a case-by-case basis, taking into account multiple factors including, but not limited to, product type, MoA, associated risk, and phases of development. This presentation provides an overview of regulatory expectations regarding potency assays and discusses several case studies that highlight some of the relevant issues commonly seen in the regulatory submissions.

10:10 Bioassay and Immunoassay Assessment of an ADC

Jeffrey_GlennJeffrey Glenn, Ph.D., Associate Director, Analytical Development, Bristol-Myers Squibb

Potency of ADCs can be measured using ELISA and cell-based assays. Both methodologies are sensitive to changes in antibody-drug ratios and modifications to the CDR-regions of the molecule. By employing both ELISA and cell-based bioassay a complete picture of the ADC can be obtained. This talk will present data from both types of assays, and describe how they can be used to evaluate manufacturing and lot-to-lot changes.

10:40 Coffee Break in the Exhibit Hall with Poster Viewing


11:10 New MS- and LC/MS- Based Approaches to Characterization of Highly Heterogeneous Biopharmaceuticals

Igor_KaltashovIgor A. Kaltashov, Ph.D., Professor and Graduate Program Director, Department of Chemistry, University of Massachusetts-Amherst

Characterization of biopharmaceuticals focuses on their structure at a variety of levels, and the heterogeneity inherent to many protein-based therapeutics makes it a truly challenging task. This presentation will feature several analytical methods recently developed in our laboratory that allow this task to be accomplished for a variety of highly heterogeneous targets including stressed biopharmaceuticals, PEGylated proteins and other protein conjugates. The main focus will be on size exclusion and ion exchange chromatography with on-line detection by native MS and top-down MS/MS.

11:40 Applications of LC-MS Towards the Understanding and Characterization of High Molecular Weight Variants in the Development of Biotherapeutics

Richard Ludwig, Ph.D., Principal Scientist, Molecular Analytics and Development, Bristol-Myers Squibb

LC MS has traditionally played a significant role in the characterization of biotherapeutics. This role has until recently been limited to assessment of the primary structure of proteins. Recent advancements in LC-MS techniques have enabled the use of LC-MS as a means of investigating higher order interactions. The application of hydrogen deuterium exchange as a means of characterizing a high molecular weight species within a developmental monoclonal antibody biotherapeutic is described.

12:10 pm Presentation to be Announced


12:40 Luncheon Presentation (Sponsorship Opportunity Available) or Lunch on Your Own


1:55 Chairperson’s Remarks

Jeffrey W. Hudgens, Ph.D., Research Chemist, Biomolecular Measurement Division, Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology

2:00 Comparability and Monitoring N-Linked Oligosaccharides Using HPLC and Mass Spectrometry

Xiaoyang Zheng, Ph.D., Senior Scientist, Analytical Development, Sanofi US

One of the most important structural features of recombinant therapeutics is the N-linked oligosaccharide profile. These profiles impact recombinant therapeutics in a wide range of ways by affecting their pharmacokinetics, potency, safety, and stability. Oligosaccharide profiling using HPLC, sequential digestion with exoglycosidases, and mass spectrometric analysis will be presented. And the potential impact of oligosaccharides on function and stability of the glycoprotein therapeutics will be discussed in this talk.

2:30 Fc Effector Functions, FcRn and Their Impact on PK/PD of Antibodies, and Potential Implications for Comparability and Biosimilarity

 Qinwei_ZhouQinwei Zhou, Ph.D., Vice President, BioAnalytical Science, Eli Lilly and Company  

Monoclonal antibodies have demonstrated safety and efficacy with favorable clinical response, resulting in many commercial products on the market. Several factors determine the efficacy, including target specificity through Fab, effector functions and half-life through Fc. The regulatory expectations and requirements for detailed characterization of Fc’s part are increasing. This presentation will share our strategy.

3:00 Strategies for the Control of Glycosylation of a Chimeric Human-Llama Antibody

Michael_ButlerMichael Butler, Ph.D., Scientific Director, MabNet, Distinguished Professor, Department of Microbiology, University of Manitoba

The glycoform profile of a chimeric monoclonal antibody can be controlled to pre-defined levels- galactosylation, fucosylation and sialylation. These common variants of the conserved Fc glycan have a significant effect on the functional properties of the molecule that may affect therapeutic efficacy. The presentation will review the bioprocess parameters that can be controlled in order to minimize batch to batch variation of glycosylation and obtain a desired glycan profile.

3:30 Sponsored Presentation (Opportunity Available)

4:00 Refreshment Break in the Exhibit Hall with Poster Viewing

4:30 Sequence Variants and Sequence Variant Analysis in Biotherapeutic Proteins

Oleg V. Borisov, Ph.D., Director, Analytical Development, Novavax

A “sequence variant” is a surrogate term covering any unintentional amino acid substitutions, omissions, or insertions during protein biosynthesis. The occurrence of sequence variants contributes to heterogeneity of recombinant protein therapeutics. Establishing a sequence variant profile of a biotherapeutic product is essential in providing proof of its structure, its manufacturing consistency, and the stability of the producing cell line. This talk will discuss analytical methodologies for detecting sequence variants in biotherapeutic proteins, and will discuss the results of an intra-laboratory study for sequence variants analysis in the NIST monoclonal antibody (mAb) by peptide mapping.

5:00 Assessment of Critical Quality Attributes in Biopharmaceuticals and Their Impact on Different Stages of Product Development

Robert Mayer, Ph.D., Scientist, Characterization Group, Sandoz GmbH

The adequate control of CQAs at different stages of the manufacturing process is essential, as they can directly be linked to clinical safety and efficacy. Other elements like process risk assessments or drug substance and drug product control strategies are being established based on the CQA assessments. This talk will focus on the identification and classifications of quality attributes according to their criticalities based on the available knowledge and (un)certainty thereof. The impact of CQAs on different stages of the product development will be discussed.

5:30 Networking Reception in the Exhibit Hall with Poster Viewing

6:30 Close of Part Two: Advances in Characterization Methods and Approaches - Stay on for Part Three: Comparability and Biosimilarity  

Day 1 | Day 2 | Download Brochure | Speaker Bios