Cambridge Healthtech Institute’s 2nd Annual
Characterization of Biotherapeutics
Improving Prediction, Screening and Characterization of New Biologics
January 18-19, 2016

New biotherapeutics formats are flooding the discovery and development pipelines and with this comes an increasing need for better and faster characterization tools and strategies, improved biomolecular and biophysical assays for the new biotherapeutics. The 2nd Annual Characterization of Biotherapeutics conference presents new tools, strategies and high-level case studies on analytical development and characterization of mAbs, ADCs, bispecifics and other novel protein formats. We are looking for cutting-edge tools, research findings and unpublished data to be featured at this forum.

We invite you to present a poster and join colleagues in this discussion of the key challenges and solutions improving prediction, screening and characterization of new biologics.

Final Agenda

MONDAY, JANUARY 18

7:30 am Conference Registration and Morning Coffee

EMERGING TRENDS IN BIOANALYTICAL CHARACTERIZATION

9:00 Chairperson’s Opening Remarks

Atul Saluja, Ph.D., Senior Research Investigator II, Drug Product Science & Technology, Bristol-Myers Squibb

9:50 New USP Chapter <212> Oligosaccharide Analysis for N-glycan Analysis

Edith Chang, Ph.D., Scientific Liaison, Biologics and Biotechnology, US Pharmacopeia

The United States Pharmacopeia publishes a new General Chapter to provide qualitative analysis of glycosylation through profiling of released N-linked oligosaccharides (or N-glycans). This chapter consists of validated analytical procedures and performance criteria. Furthermore, four reference standards (RSs) have been developed to assess the system suitability for the analytical procedures. Details of this chapter and the applications of RSs are presented.

10:20 Coffee Break

PROPERTY-BASED DEVELOPMENT, CHARACTERIZATION AND DEVELOPABILITY ASSESSMENT

10:45 Conjugation and Aggregation Aspects of an Antibody-Drug Conjugate: The Intimate Link between Drug Substance Process and Drug Product Quality

Atul Saluja, Ph.D., Senior Research Investigator II, Drug Product Science & Technology, Bristol-Myers Squibb

Given the chemical complexity of an antibody-drug conjugate (ADC), it is critical to understand drug related factors that can influence its physicochemical stability during manufacturing as well as storage. This presentation will explore the impact of conjugation and drug loading on physicochemical stability for a model ADC. Learnings should be applicable to current and future ADCs for aligning drug substance and drug product development activities.

11:15 Use of a Slope Measurement Method Employing Variable Pathlength UV-Vis Technology to Determine Antibody Concentration and Payload (Drug) To Antibody Ratio (DAR) in ADCs

Sonia Taktak, Ph.D., Analytical Scientist III, Analytical and Pharmaceutical Sciences, ImmunoGen, Inc.

The traditional approach to measuring antibody concentration and DAR in ADCs is based on fixed pathlength measurement by UV-Vis, which requires use of dilution factors that are time consuming and prone to error. In this presentation, we will review results of our evaluation of a new approach that uses Solo VPE variable pathlength technology, which does not require dilution of sample, and discuss its applicability for use in ADC manufacturing.

11:45 In silico Prediction of Tryptophan and Methionine Oxidation

Alexander Jarasch, Ph.D., Bioinformatics Scientist, Pharma Research and Early Development, Therapeutic Modalities, Roche Innovation Center Penzberg, Roche

Developability assessment of therapeutic antibodies during the selection process is a crucial step of today’s drug development and helps to identify stable and manufacturable drug candidates. A frequent developability issue – besides Asp and Asn deamidation/isomerization-is the oxidation of Trp and Met. Here we present an in silico machine-learning algorithm which predicts oxidation by combining experimental mass spectrometry data with calculated structural features in antibodies.

12:15 pm Applied Developability: Finding the One in 1,000 Successful Candidate

Georg Blaser, Ph.D., Senior Group Leader, Cell and Molecular Biology, Applied Protein Services, Lonza Biologics plc

Only one in every 1,000 preclinical candidates reaches the commercial market. This presentation showcases how applied developability such as Lonza's aggregation, T cell epitope prediction and developability assessment tools can help to identify and reduce candidates at potential risk of failure during early discovery and development phases. This presentation will also explain how protein engineering can be used to address these potential sequence liabilities. Case studies will be shown that exemplify the power of these methods.

12:45 Session Break

1:00 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

TOOLS TO SUPPORT COMPARABILITY ASSESSMENT, CONTROL STRATEGY AND CRITICAL QUALITY ATTRIBUTES

2:00 Chairperson’s Remarks

Shawn Russell, Ph.D., Associate Director, Process Development, Five Prime Therapeutics

2:05 Mass Spec Application in Analytical Comparability and Biosimilarity

Yan-Hui Liu, Ph.D., Principal Scientist, Sterile Product and Analytical Development, Biological Development, Merck

2:30 Characterization of Product Variants Observed During Pre-Comparability Assessment of a Monoclonal Antibody

Shawn Russell, Ph.D., Associate Director, Process Development, Five Prime Therapeutics

Product variants were noted in protein produced from two cell line candidates during a comparability assessment. Protein from the first candidate exhibited additional peaks by IEX and an extra band by SDS-PAGE. From the second candidate, we observed an additional peak by SEC. Characterization of the product variants showed the first candidate was not appropriate to move forward. The second candidate product variant was identified and deemed comparable after further purification.

2:55 High-Throughput Analytical Technologies for Accelerated Bioprocess Development and Continuous Processing

Jun Hyuk Heo, Analytical Scientist, Bioprocess Technology and Expression (BTE), Merck Co., Inc.

With implementation of high throughput process development and continuous processing tools, real-time and ht analytics are desired to support the development. PATROL UPLC was used for monitoring aggregates in perfusion bioreactor and near-IR based system was used to monitor and feedback glucose in real time. Microfluidic immunoassay system was used for quantification of protein analytes. The real-time monitoring by PATROL and near-IR system and fast turnaround time by microfluidic immunoassay system were proven to be highly effective for rapid bioprocess development.

3:20 Reducing the N-terminal Heterogeneity of a Monoclonal Antibody using a Combination of Enzymes

Liangyi Zhang, Ph.D., Senior Scientist II, Process Sciences, Abbvie

Recombinant monoclonal antibodies (mAb) often exhibit N-terminal heterogeneity resulting from incomplete signal peptide cleavage, enzymatic processing and/or chemical modification of N-terminal amino acids. Convoluted with the amino acid degradation, the N-terminal variants lead to complex charge variant profile, making analytical characterization and stability monitoring more difficult. We have used a combination of enzymes to reduce the N-terminal and C-terminal heterogeneity of a mAb that can greatly simplify the CEX charge variant profile. It thus enabled better characterization and monitoring of other charge variants that are related to amino acid degradation.

3:35 Sponsored Presentation (Opportunity Available)

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

4:30 Scouting for Chromophores in Protein Therapeutics: A Systematic Approach

Hangtian Song, Ph.D., Senior Scientist I, Global Manufacturing & Supply, Biologics Development & Operations, Bristol-Meyer Squibb

CHARACTERIZATION OF BIODISTRIBUTION AND IN VIVO FATE OF BIOLOGICS

4:55 Ocular Pharmacology: Probing Biotherapeutic Systemic Effects and Stability in Ocular Matrices

Kelly Loyet, Ph.D., Scientist, Biochemical and Cellular Pharmacology, Research, Genentech, Inc.

Since the frequency of ocular biotherapeutic dosing is often monthly or longer, it is important that the biotherapeutic maintains ocular stability between dosing. This study examines both in vitro and in vivo ocular stability of a biotherapeutic. Immunoassay as well as mass spectrometry and SPR methods were used to probe attributes of the biotherapeutic over time. Long-term in vitro ocular stability was shown to translate to in vivo ocular stability.

5:20 Influence of the Antibody Variable Domain Charge Distribution on In-Vivo Fate and Pharmacokinetics

Hubert Kettenberger, Ph.D., Senior Principal Scientist, Roche Pharma Research and Early Development (pRED), Roche Innovation Center Penzberg

The interaction of monoclonal antibodies (mAbs) with the neonatal Fc receptor (FcRn) is essential for their long serum half-life. Comparing a series of mAbs with identical Fc domains, we identified a contribution of the variable region to FcRn binding and pharmacokinetics which correlates with the charge distribution in these variable domains. These findings can guide the design and selection of mAbs with long serum half-life.

6:30-7:45 Welcome Reception in the Exhibit Hall with Poster Viewing

7:45 Close of Day

TUESDAY, JANUARY 19

8:00 am Conference Registration and Morning Coffee

CHARACTERIZATION OF BISPECIFIC ANTIBODIES AND NEW BIOTHERAPEUTICS

8:30 Chairperson’s Remarks

Michael Hunter, Ph.D., Associate Director, Protein Sciences, Janssen Biotechnology

8:35 Development and Characterization of Novel Antibody Formats

Melissa Geddie, Ph.D., Principal Scientist, Discovery, Merrimack Pharma

Multispecific antibodies and antibody-like molecules broaden the therapeutic application of IgGs, but can be challenging to engineer and manufacture. Using a network biology approach to identify key design parameters, we have engineered novel formats for specific biological targeting. We then use rapid design cycles followed by high-throughput characterization of these formats to select for potential therapeutic candidates with robust pharmaceutical properties.

9:05 Purity Analysis of Bispecific Antibodies by Affinity Capillary Electrophoresis

Kathir Muthusamy, Ph.D., Staff Scientist, Regeneron Pharmaceuticals, Inc.

Despite wide therapeutic applications for bispecific antibodies (bsAbs), challenges associated with manufacture and purity analysis prevail. Co-expression of bsAbs with common light chain minimizes the number of homodimers (from 10 to 3) that are subsequently removed during purification. Conventional purity analysis methods are inadequate due to analogous physiochemical properties of bsAbs and homodimers. To address this challenge, a robust and powerful Capillary Zone Electrophoresis (CE) method combined with affinity CE has been developed.

9:35 Cutting Edge Vibrational Spectroscopy for Protein Therapeutics

Rina K. Dukor, Ph.D., President, CEO, BioTools, Inc.

The use of FT-IR spectroscopy as a probe of secondary structure is now widespread throughout the biopharmaceutical industry. More recently, ROA (Raman Optical Activity) has been shown to indicate differences when none are observed with any other spectroscopic technique. In this presentation, we will discuss advances in four forms of vibrational spectroscopy as applied to structural studies of proteins.

9:50 Coffee Break in the Exhibit Hall with Poster Viewing

11:00 Developability Risks that Are Particular to Bispecific Antibodies

Michael Hunter, Ph.D., Associate Director, Protein Sciences, Janssen Biotechnology

Multispecific antibodies deliver novel molecules that can access differentiated mechanisms of action. Preparations of bispecific antibodies can affect various criteria for manufacturability including protein integrity, structure-function, and serum compatibility. A review of the risks and mitigation will be provided.

11:30 Challenges and Chances for Bioanalysis of Bispecific Antibodies

Kay Stubenrauch, Ph.D., Senior Principal Scientist, Pharmaceutical Sciences, Roche/Genentech, pRED

While the emerging class of bispecific mAbs is characterized by the diversity of molecular constructs, it has in common two sets of different complementarity determining regions (CDRs). Bispecific antibodies also provide new challenges and chances in soluble target assays and immmunogenicity testing. A good understanding of the target biology and of the nature of the bispecific antibody combined with timely availability of specific assay reagents will facilitate more complex bioanalytical methods for analysis of bispecific antibodies.

12:00 pm Sponsored Presentation (Opportunity Available)

12:30 Session Break

12:45 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:45 Close of Conference