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Analytical Methods & Technologies

Analytical Methods & Technologies

Analytical Methods & Technologies

Monday, March 30, 2015 - Thursday, April 2, 2015

A must-attend track for scientists seeking novel analytical methods for accurate characterization of biologics. From comparability and characterization strategies to QbD concepts to aggregation and subvisible particle analysis, get the tools and applicable knowledge you need to enhance your operation's process and product quality.

» Download Agenda | Register Now

Session Highlights

  • Keynote or Featured Presentation
  • Case Study
  • New Data
  • Panel Discussion
  • Speaker Interview

Analytical Methods & Technologies

Monday, March 30, 2015

7:00
Registration and Coffee

Keynote Presentations

8:00
Chairperson's Opening Remarks
Günter Jagschies, Ph.D., Senior Director, Strategic Customer Relations, Biotechnologies R&D, GE Healthcare Life Sciences, Sweden

8:15
Presentation TBA

9:00
Dana Andersen, Ph.D. Challenges and Opportunities in Pharmaceutical Development
Increasing needs to improve the efficiency of development together with new opportunities such as Breakthrough Therapy designation, will require novel approaches and innovations to be successful. Specific examples and opportunities will be discussed focusing on drug product development but many of which are more broadly applicable as well.
Dana C. Andersen, Ph.D., Senior Director, Pharmaceutical Development, U.S. Biologics Technical Development, Genentech - a Member of the Roche Group

9:45
Networking Refreshment Break in Poster and Exhibit Hall

10:25
Chairperson's Remarks
Parastoo Azadi, Ph.D., Technical Director, Complex Carbohydrate Research Center, University of Georgia

Formulation & Delivery Developments

10:30
Developing Drug/Device Combination Products for Oral Inhalation
Developing drug/device combination products requires early integration of formulation and device technologies. For inhalation products, dry powder formulations and inhalation devices are intrinsically linked and product development challenges can be simplified when these efforts are combined at an early stage. This presentation will address these challenges with case studies of inhaled drug/device combinations products.
Andrea Leone-Bay, Vice President, Pharmaceutical R&D, MannKind

Comparability Strategies for Biosimilars and Other Biotechnology Products

11:00
Building Mass Spectral Libraries for the NISTmAb Reference Material
NIST is developing a curated and fully evaluated reference mass spectral (MS) library of all identifiable components, including peptides, glycans, and glycopeptides, derived from the digestion of its future monoclonal antibody reference material, NISTmAb. This library is an extension of the existing NIST tandem MS libraries of peptides and glycans and includes software tools for spectral searching and scoring.
M. Lorna De Leoz, Ph.D., NIST Mass Spectrometry Data Center, National Institute of Standards and Technology

11:30
Case Studies on Comparability Assessment of Biological Drug Products
Overview of Drug Product changes that necessitate demonstration of comparability and general approaches for appropriate comparison of Drug Product quality attributes of the pre-change to post-change will be discussed. Accelerated and stress stability studies are often useful tools for comparison of degradation rates and degradation profiles of pre-change to post-change Drug Products. However, stress comparability assessment of biological Drug Products has its own challenges. Case studies on stress comparability assessment of biological Drug Products during manufacturing site transfers and process changes will be presented.
Camellia Zamiri, Ph.D., Scientist, Genentech, a Member of the Roche Group

Technology Workshop Presentations

12:05
Single-Use Fermentation: Understanding Process Economy and Process Performance
The entry of single-use bioprocessing and the benefits that come with disposables have generated interest in the microbial community. This talk will present results from a process economy comparison of fermentation scenarios based on stainless steel and single-use equipment. Additionally, data will be shown from a wet-work study of an E.coli domain antibody production using stainless steel and disposable equipment.
Kenneth P. Clapp, M.S., Senior Product Manager, Bioreactors & Fermentors, GE Healthcare Life Sciences / Xcellerex

Fit For Purpose – Design and Implementation of Single-Use Bioprocessing Plants
Flexibility and speed are important factors for clinical manufacturing. Most of the biopharmaceutical manufacturers have initiated programs to investigate alternative ways to produce clinical material in order to improve these two requirements. However there are essential matters which have to be considered during implementation. In this contribution case studies of successfully executed fast track projects for Phase I-III material will be shown.
Dr. Thorsten Peuker, VP Integrated Solutions, Sartorius Stedim Biotech

Using Process and Activity to Drive Clone Selection
During a manufacturing CHO stable cell line development project, it was observed that the product being made by the CHO clones displayed a significant reduced specific activity when compared to a reference standard. This case study highlights how the lack of activity was identified and how the development of a cell culture process enabled the identification of high productivity clones.
Oren E. Beske, Ph.D., COO, Aragen Bioscience, Inc

12:35
Networking Luncheon in Poster and Exhibit Hall

1:40
Chairperson's Remarks
John P. Marino, Ph.D., Leader, Biomolecular Structure & Function Group, NIST

Comparability Strategies for Biosimilars and Other Biotechnology Products (continued)

1:45
Tina Morris The Role of Pharmacopeial Standards in the Context of Biosimilars
Tina Morris, Vice President, Biologics & Biotechnology, United States Pharmacopeia

2:15
Application of NMR Spectral Fingerprinting for Structure Assessment of Monoclonal Antibody Therapeutics
High-resolution NMR methods can provide spectral 'fingerprints' of the higher order structure of protein therapeutics at atomic resolution. This presentation will report on the application of NMR methods for obtaining NMR 'fingerprints' of monoclonal antibody therapeutics and how these spectral 'fingerprints' can be used to establish consistency in drug manufacturing and for comparing a biosimilar to an innovator reference product.
John P. Marino, Ph.D., Leader, Biomolecular Structure & Function Group, NIST

2:45
Success and Lessons Learned in Characterization and Comparability
Moderator: Parastoo Azadi, Ph.D., University of Georgia
Panelists:
M. Lorna De Leoz, Ph.D., National Institute of Standards and Technology
Camellia Zamiri, Ph.D., Genentech, a Member of the Roche Group
John P. Marino, Ph.D., NIST
Tina Morris, United States Pharmacopeia

3:15
Networking Refreshment Break in Poster and Exhibit Hall

Improving and Controlling Process and Product Quality / Impact of Cell Culture on Critical Quality Attributes

4:00
The Production of Antibodies as Single Glycoformsor with a Restricted Glycosylation Profile
The glycoform profile of a monoclonal antibody (Mab) determines many functional properties that affect therapeutic efficacy. The glycan profile can depend upon the producer cell line, the growth media, the culture conditions as well as the Mab structure. The presentation will review bioprocess parameters that can be controlled to affect this glycosylation profile. Strategies will also be discussed to produce Mabs with pre-defined glycan structures by controlling the culture conditions or by enzymically re-modelling glycans during the purification process.
Michael Butler, Ph.D., Professor of Microbiology, University of Manitoba, Canada

4:30
Challenges in Comparability Studies of CarbohydrateContaining Biosimilars
The analysis of the glycoconjuagtes has become important for all comparability studies and in the quality control of therapeutic recombinant glycoproteins and polysaccharides. We will describe challenges in chromatography and mass spectrometry methods and procedures that are used for polysaccharide and glycoprotein biosimilar analysis in order to obtain regulatory approval. Details will be given on the pros and cons of certain methods from regulatory stand point. We will discuss procedures necessary for the complete structural elucidation of heparin, low molecular weight heparin and antibody products. Challenges in comparability study of low molecular heparin will be discussed. Examples of the methodologies will include data needed in different phases and currents analysis of biosimilars.
Parastoo Azadi, Ph.D., Technical Director, Complex Carbohydrate Research Center, University of Georgia

5:00
Advances and Challenges in the Analytical Characterization of Biosimilar Products
At the core of every biopharmaceutical development program is the challenge to successfully produce active and high quality protein, however for biosimilars additional challenges arise in demonstrating the comprehensive analytical comparability to the reference product. Utilizing our technology, Pfenex has rapidly accelerated the development of a biosimilar candidate, demonstrated analytical biosimilarity and progressed the product into clinical trials.
Jeff Allen, Ph.D., Director of Protein Sciences, Pfenex Inc.

5:30
Welcome to the BDP Big Top!
Grab your popcorn and peanuts as you make your way through the exhibit hall for your front row look at the most amazing display of biopharmaceutical technologies and services all under the BDP Big Top!

Tuesday, March 31, 2015

7:30
Registration and Coffee

8:00
Chairperson's Remarks
Alex Lazar, Ph.D., Head of Analytical and Pharmaceutical Sciences, ImmunoGen, Inc.

Analytical Characterization for ADCs

8:15
Yan Chen, Ph.D. Overcoming Analytical Challenges during Development and Production of ADCs
Overview of Drug Product changes that necessitate demonstration of comparability and general approaches for appropriate comparison of Drug Product quality attributes of the pre-change to post-change will be discussed. Accelerated and stress stability studies are often useful tools for comparison of degradation rates and degradation profiles of pre-change to post-change Drug Products. However, stress comparability assessment of biological Drug Products has its own challenges. Case studies on stress comparability assessment of biological Drug Products during manufacturing site transfers and process changes will be presented.
Yan Chen, Ph.D., Technical Development Senior Scientist, Protein Analytical Chemistry, Genentech, Inc.

8:45
Real-Time Monitoring of ADC Conjugation Process
Antibody Drug Conjugates have process development and manufacturing challenges in part due to the uniqueness of combining a tumor targeting antibody with a potent small molecule cytotoxic agent. This presentation shows the application of real time monitoring of ADC conjugation reactions to process development and potential usefulness of real time monitoring during scale up and manufacture.
Ian Schwartz, M.S., Senior Engineer, ADC Process Development, Agensys, an affiliate of Astellas

9:15
Analytical Development and Characterization Tools(3-D Structure of mAb/ADCs, Mass Spec)
Oscar Salas-Solano, Ph.D., Senior Director of Analytical Sciences, Seattle Genetics, Inc.

9:45
Networking Refreshment Break in Poster and Exhibit Hall

10:25
Chairperson's Remarks
Thomas A. Little, Ph.D., President, Thomas A. Little Consulting

QbD/CQA Concepts from Development to Submission

10:30
Essentials in Quality by Design for Modern Biologics Development
QbD is a systematic approach to development that begins with predefined development objectives and emphasizes product and process understanding and process control, based on sound science, data based decision making and quality risk management. Quality by Design as introduced by the FDA and EU brings modern drug development methodologies to CMC teams working on biologics, pharmaceuticals and vaccines. The presentation will discuss the application of QbD principles in the development, submission and manufacturing of drug product and drug substance. Most CMC development teams globally have little to no experience in generation of design space, selection of PAR/NOR ranges and preparing for Stage I Validation documentation and submission. This presentation will cover basic and advanced principles for the practical application of QbD in every phase of product development and control.
Thomas A. Little, Ph.D., President, Thomas A. Little Consulting

11:00
The Role and Design of Forced Degradation Studies in Biologics Development
Xiaoyu Chen, Section Leader, Late Stage Analytics, Merck

11:30
Successfully Applying QbD Concepts to Develop a Robust Lyophilized Protein Therapeutic Product
This case study demonstrates how to apply Quality-by-Design (QbD) concepts to develop a robust lyophilized dosage form for an inherently unstable protein therapeutic. The presentation will walk through design and execution processes: risk assessment, analytical method readiness, multivariate statistical design, acquisition of representative data, and data crunching in order to propose a sound product formulation in the context of designated container closure systems.
Kevin Zen Ph.D., Senior Manager, Biologics Development, Allergan

Technology Workshop Presentations

12:05
New Low Shear Force Single-Use Pump System enables High Cell Viabilities, Protein Activities and a Safe, Pulsation Free Conveyance of Sensitive Biotech Media
Cells and proteins used in today's Biotech processing can be very sensitive to external mechanical forces such as shear stress. Commonly used technologies as peristaltic pumps can cause severe influence on cell viability or protein activities. This workshop introduces a new pump technology applying magnetic levitation for a safe, pulsation free conveyance of sensitive biotech media. Current research at the ZHAW University Zurich reveals astonishing results comparing the influence of different pump systems to cells and proteins which will be shared during the presentation.
Wolfgang Dornfeld, MSc, BSc, MBA, VP Field Operations, Levitronix GmbH
Ina Dittler, MSc, Research Asst, School of Life Sciences & Facility Management, Zurich University of Applied Sciences

Agile Development of a Biobetter ADC: A Case Study
Antibody-drug conjugates (ADC's) represent a promising therapeutic approach combining the antigen-targeting specificity of monoclonal antibodies (mAbs) with the cytotoxic potency of chemotherapeutic drugs including the essential linkers. Creating a vertical hierarchy will allow agile benefits such as high levels of product customization, stringent quality control, shorter development and production spans, and economies of scale. EirGenix and Formosa Laboratories have formed an alliance that provide for all components of ADC drug development. Through the implementation of this agile hierarchy they have developed a unique Bio-better.
Molly McGlaughlin, VP Business Development, EirGenix

12:35
Networking Luncheon in Poster and Exhibit Hall

1:40
Chairperson's Remarks
Mark Emanuele, Process Engineer, Genzyme

Aggregation and Subvisible Particle Analysis in Protein Drug Products

1:45
Resolution of Heterogeneous Charged Antibody Aggregates via Multimodal Chromatography:Comparison to Conventional Approaches
Clearance of aggregates during protein purification is increasingly paramount as protein aggregates represent one of the major impurities in biopharmaceutical products. Aggregates, especially dimer species, represent a significant challenge for purification processing since aggregate separation coupled with high purity protein recovery can be difficult to accomplish. Biochemical characterization of the aggregate species from the hydrophobic interaction and cation exchange chromatography elution peaks revealed two different charged populations, i.e. heterogeneous charged aggregates, which led to further challenges for chromatographic removal. This paper compares multimodal versus conventional cation exchange or hydrophobic chromatography methodologies to remove heterogeneous aggregates. A full, mixed level factorial design of experiment strategy together with high throughput experimentation was employed to rapidly evaluate chromatographic parameters such as pH, conductivity, and loading. A variety of operating conditions were identified for the multimodal chromatography step, which lead to effective removal of two different charged populations of aggregate species. This multimodal chromatography step was incorporated into a monoclonal antibody purification process and successfully implemented at commercial manufacturing scale.
Rebecca A. Chmielowski, MSD, Biologics Process Development, Process Development and Engineering, Merck

2:15
Analyzing Subvisible Particles in Protein Drug Products: A Comparison of Dynamic Light Scattering (DLS) and Resonant Mass Measurement (RMM)
Aggregation is common in protein drug manufacture, and while the effects of protein particulates are under investigation, many techniques applicable for their characterization have been recently developed. Among the methods available to characterize and quantify protein aggregates, none is applicable over the full size range and different methods often give conflicting results. The studies presented here compare two such methods: dynamic light scattering (DLS) and resonant mass measurement (RMM). The performance of each method was first characterized using polystyrene particle size standards (20, 60, 100, 200, 400, and 1,000 nm) over a range of concentrations. Standard particles were measured both singly and in binary mixtures containing 20 nm particles at a fixed concentration (10(14) particles/mL) and various concentrations of one of the other particle sizes (i.e., 60, 100, 200, 400, or 1,000 nm). DLS and RMM were then used to detect unknown aggregate content in stressed samples of IgG. Both instruments were shown to have a working range that depends on particle size and concentration. In binary mixtures and polydisperse solutions, DLS was able to resolve two species in a manner dependent on both concentration and particle size. RMM was able to resolve particles above 200 nm (150 nm for protein) at concentrations below 10(9) particles/mL. In addition, dilution was evaluated as a technique to confirm and quantify the number of particles in solution.
Jainik Piyushkumar Panchal, Post Doctoral Fellow, Purdue University

2:45
Networking Refreshment Break in Poster and Exhibit Hall

Analytical Characterization Methods for Critical Quality Attributes

3:30
Process Characterization of Vaccine Candidates: Where Do We Start?
When we speak of process characterization, we often think of a process in their late phase of clinical manufacture and qualification. Usually, there is a desire to characterize "everything" without really going a systemic methodology of performing characterization. In this talk, we will be looking at some of the ICH guidelines and how we use those guidelines to appropriately stage process characterization. The discussion would cover examples of process characterizing microbial expressed vaccine antigens. The process starts with performing a process risk assessment, performing small-scale model, characterizing unit operations to obtain operating parameters to control your process to meet critical quality attributes.
Eric Chojnicki, Ph.D., Director CMC, Biologics Development, Allergan

4:00
Platform for Media and Bioreactor Nutrient Analysis
A platform for media nutrient characterization was established at the contract research organizations (CROs) and Allston MSAT Laboratory. Analytical methods of Ultra Pressure Liquid Chromatography Triple Quadrapole Mass Spectrometry (UPLC/MS/MS) and Inductively Coupled Plasma Mass Spectrometry (ICP/MS), as well as Blood Gas Analyzer (BGA), were used to analyze formulated media and bioreactor retains for amino acids, vitamins, trace elements and sugars. Cell culture media samples (pre- and Post a 0.1 µm filter) were collected over a six-month period and analyzed to determine if the nutrient concentrations were consistent and if they were in agreement with theoretical target values. In addition, a comparison between pre- and post-filter samples was performed to identify if nutrient loss was occurring during the filtration process. Perfusion bioreactor samples (spent media) were collected from the growth, transition and harvest phases to understand the nutrient profiles. The study results show the formulated media is consistently prepared and is at the theoretical concentrations, with the exception of two components. The study results also showed loss after filtration for one component. The bioreactor nutrient profiles show a decrease or depletion of several nutrients, while an increase of other nutrients were observed.
Mark Emanuele, Process Engineer, Genzyme

4:30
James Swartz, Ph.D. Engineering Novel Proteins and Protein Assemblies as Innovative Therapeutics, Vaccines, and DrugDelivery Vehicles
The exciting emergence of bispecific antibodies and antibody drug conjugates as new, important classes of protein therapeutics paves the way for developing new families of novel, relatively complex therapeutics. This includes protein assemblies containing multiple copies either of the same or of different proteins as well as protein complexes displaying and/or containing other types of molecules. It is clear, however, that the assembled product form must be consistent and must remain stable during storage and administration. We are now developing modular vaccine forms designed to mimic viruses while also triggering immune stimulation mechanisms evolved to fight bacterial infections. A novel influenza vaccine candidate is comprised of a precisely assembled virus-like particle (VLP) composed of 240 subunits to which multiple copies of a trimeric hemagglutinin stem antigen have been attached with a precise presentation orientation. Finally, a DNA oligomer is co-attached to stimulate an innate immunity thereby maximizing the protective response to the antigen. In this talk, I will describe the precautions taken during the development of this new entity to ensure that a uniform and stable product results. Multiple mutations as well as careful process development were required to enhance folding and assembly of both the VLP and antigen monomers as well as to stabilize the multimeric assemblies. I will then describe how the same approach is being used to develop multi-functional drug delivery vehicles using specially engineered VLPs.
James R. Swartz, Ph.D., James H. Clark Professor, School of Engineering, and Professor of Chemical Engineering and of Bioengineering, Stanford University

5:00
Close of Day Two

5:30
BDP Week Beach Party
Back by Popular Demand!
Shimmering views of the Pacific Ocean will be the back drop at this fun, casual networking reception. You'll enjoy beach-themed food, drink and music in this unparalleled coastal setting as you network with speakers, attendees and exhibitors. Free for All Registered Attendees - RSVP required.
Co-Sponsored by: and

Session Highlights

  • Keynote or Featured Presentation
  • Case Study
  • New Data
  • Panel Discussion

Analytical Methods & Technologies

Wednesday, April 1, 2015

7:30
Registration and Coffee

Keynote Presentations

8:00
Chairperson's Remarks
Uwe Gottschalk, Ph.D., Chief Technology Officer, Pharma Biotech, Lonza, Switzerland

8:15
Tae Han Kim Manufacturing in a Global Market Place
As the CEO of leading contract manufacturing organization - Samsung BioLogics, Dr. TH Kim presents the implication of manufacturing segment in the whole business value chain of biopharmaceutical industry. Beginning with the trend forecast of growing demand and supply of manufacturing capacity, he talks about the limitation of in-house manufacturing and a solution to manufacturing optimization in global market place.
Tae Han Kim, Ph.D., President and CEO, Samsung BioLogics

8:45
Hitto Kaufmann, Ph.D. Innovation in Biologics Manufacturing
Hitto Kaufmann, Ph.D., Vice President, Technology and Innovation, Sanofi Biologics, Germany

9:45
Networking Refreshment Break in Poster and Exhibit Hall

10:25
Chairperson's Remarks
Fengqiang Wang, Ph.D., Principal Scientist, Sterile Product and Analytical Development, Merck

Quantitation and Control Strategies for HCPs

10:30
Antigen Excess and Avoiding HCP Quantitation Errors
Immunoassays are typically used to monitor residual host cell protein (HCP) clearance from biopharmaceuticals. Antibodies to a complex mixture of total HCPs are generated for this purpose. One quantification error potentially occurs if a particular protein co-purifies with product and is present in excess of the available anti-HCP antibodies in the assay ("antigen excess"). We have explored different assay formats to monitor and address the antigen excess phenomenon.
Sara Parker, Ph.D., Senior Manager Analytical Operations, Genentech, a Member of the Roche Group, Inc.

11:00
Analytical Challenges and Solutions for In-House HCPs Assay Development, Case Studies from HCPs Antigen Selection to Anti-HCPs Reagents Qualification
ELISA is the most commonly used method for monitoring HCPs clearance during biologics production. The performance of HCP ELISA is highly relied on the anti-HCPs reagents used. This talk will cover the major analytical challenges occurred during in-house HCP assay development, from antigen selection to antibody qualification, with the focus on finding the scientific solutions to overcome these challenges.
Fengqiang Wang, Ph.D., Principal Scientist, Sterile Product and Analytical Development, Merck

11:30
Comparing a New Methacrylate Protein A Resin with Agarose Protein A-Focus on Host Cell Protein Clearance
A case study is presented whereby one of the newly developed macroporous polymethacrylate Protein A resins, Amsphere Protein A, was evaluated for binding capacity, pressure-flow characteristics, alkaline stability, and impurity clearance capabilities. As part of impurity clearance several unique wash solutions were evaluated for their ability to further improve host cell protein reduction. Results are compared to agarose Protein A.
Edward Koepf, Ph.D., Process Development Scientist, Biogen Idec

Concurrent Technology Workshop Presentations

12:05
Next Generation Technology for Continuous Chromatography
Periodic counter current chromatography (pcc) maximizes resin capacity resulting in higher productivity and smaller equipment footprint. A case study of a continuous process, which includes capture chromatography, viral inactivation and polishing chromatography, performed on the ÄKTA™ platform, including the new ÄKTA pcc system, will be presented. Comparing with a batch process, initial results indicate economic and throughput improvements while maintaining product quality.
Maria Ekblom, MSc., Senior Project Manager, Chromatography Systems, GE Healthcare

Capture Chromatography: Alternativesfor Downstream Process Development
Shelly Parra, M.S., Sr. Field Application Scientist, Thermo Fisher Scientific

How to Rapidly Enable Flexible Biomanufacturing at the Site of Your Choice
The burden of cancer and rheumatoid arthritis in a growing and ageing population is driving up the global demand for monoclonal antibodies. By attending this presentation, you will discover how the FlexFactory™ biomanufacturing platform enables to quickly increase or build production of biomolecules such as monoclonal antibodies and vaccines. Based on single-use bioprocessing equipment from cell culture to bulk product formulation, FlexFactory is a centrally automated production train that can be tailored to fit a new or existing facility within 9 to 12 months in a cost effective manner.
Vikas Gupta, Modality Leader, Bioprocess Solutions, GE Healthcare Life Sciences (invited)

12:35
Networking Luncheon in Poster and Exhibit Hall

1:40
Chairperson's Remarks
Vladimir Razinkov, Principal Scientist, Process & Product Development, Amgen, Inc.

Quantitation and Control Strategies for HCPs (continued)

1:45
Novel Impurity Challenge Study in mAb AEX Polishing Step
Bo Qi, M.S., Director, Process Development Downstream, Eli Lilly and Company

Chromatography Considerations to Combat HCPs

2:15
Membrane Filtration Can Substitute Chromatography Purification Steps for Plant-Derived and ELP-Tagged Biopharmaceutical Proteins
Chromatographic techniques are most frequently used for the purification of biopharmaceutical proteins. However, identifying an effective capture step can be a challenging task for non-antibody target proteins, especially if these are expressed at low levels or contaminated by a large number of host cell proteins (HCPs) as it is often the case for plant-derived products. In such cases, several HCPs can bind to the capture resin and reduce the effective binding capacity resulting in the necessity for large column dimensions and increasing downstream processing (DSP) costs. A selective removal of HCPs prior to the initial capture step may help to circumvent this problem and reduce costs in DSP. Here we present how membrane based pre-capture purification strategies can simplify DSP and replace chromatographic purification steps for various proteins. On the one hand, we highlight how a design of experiments approach can be used to identify conditions and membrane cut offs for the separation of plant HCPs and target proteins of different molecular masses. On the other hand, we show how membrane inverse transition cycling can be implemented into a scalable production of ELP-tagged target proteins. Both techniques can be easily adapted to other expression systems and thus may be of interested to colleagues from academia as well as from industry. This is especially true for those working with novel non-antibody products of next generation biopharmaceuticals like enzymes and vaccine candidates.
Johannes Felix Buyel, M.Sc., Institute for Molecular Biotechnology, RWTH Aachen University/Integrated Production Platforms, Fraunhofer IME, Germany

High-Throughput Methods and Strategies

2:45
Multidimensional Analysis of mAb Formulations by High Throughput Methods
Successful development of monoclonal antibodies (mAbs) depends on a thorough characterization of the molecule both before and after formulation. Characterization of a formulated mAbs, requires a rigorous assessment of the molecule's critical attributes. Critical attributes of a biotherapeutic include not only chemical stability, i.e., degradation of the molecule to form undesired modifications, but also structural stability, including the formation of aggregates and particles. Both classes of stability, chemical and structural can be assessed using a variety of characterization techniques. Recent improvements in traditional technologies, development of new methods, and advancements in computer science have created excellent innovation opportunities for high throughput technologies development. Computer-controlled and automated sample preparation and data acquisition have made those technologies amenable to a high throughput format, saving significant resources at every step of the drug development process starting with early candidate selection and continuing into clinical and commercial formulation development. In this presentation high throughput methodology for formulation analysis is described according to its applications during biopharmaceutical development of monoclonal antibodies. The characteristics obtained by high throughput methods include structural integrity, colloidal stability and chemical modifications. The power of statistical analysis, Quality by Design (QbD) and design of experiment (DOE) approaches is illustrated with several case studies.
Vladimir Razinkov, Principal Scientist, Process & Product Development, Amgen, Inc.

3:15
Networking Refreshment Break in Poster and Exhibit Hall

High-Throughput Methods and Strategies (continued)

4:00
High Throughput Determination of IgG Titer in Cell Culture via Affinity Chromatography
In Biotech industry, therapeutic IgGs are mainly produced in two expression systems: Chinese Hamster Ovarian Cell (CHO) and E. coli Cell. In order to determine harvest titer accurately for specific IgG related products, we have developed two rapid affinity chromatography methods targeting different regions on the IgG: 1) 1.5-minute Protein A chromatography method with parallel HPLC setup for IgGs from CHO and 2) 5-minute 2-dimensional Affinity(LC-Kappa®)/Reversed Phase HPLC method for IgG fragments from E. coli. In addition, automation tools used to perform high throughput sample preparation will be discussed in this presentation.
Kevin Lin, Research Associate, Genentech, a Member of the Roche Group

4:30
Assays of Higher Throughput for Efficient Measurement of Drug Potency and Impurity
Assays of higher throughput are needed for meeting the challenge of increasing demand of sample testing during drug development and production. In Biogen we have developed high throughput assays for both potency and impurity measurements. For potency assays, we use "fit-for-purpose" assay formats for testing samples from different stages of product development. For impurity assays, a case study will be presented on the development of a high sensitivity, high throughput assay for quantitation of residual host cell DNA in purification process intermediate and drug substance samples. This new method gives results comparable to our previous method, but significantly lowered the limit of quantitation and increased assay throughput.
Wei Zhang, Ph.D., Principal Scientist, Analytical Development, Biogen Idec

5:00
In-Line Separation by Capillary Electrophoresis Prior to Analysis by Top-Down Mass Spectrometry Enables Sensitive Characterization of Protein Complexes
To analyze intact proteins in the yeast Dam1 complex, we coupled capillary electrophoresis to an Orbitrap Elite mass spectrometer. We achieved a 100-fold increase in sensitivity compared to a RPLC-MS analysis of the Dam1 complex with a total loading of 2.5 ng. N-terminal processing forms of individual subunits were observed as well as their phosphorylation stoichiometry upon Mps1 kinase treatment.
Xuemei Han, Ph.D., Research Associate, The Scripps Research Institute

5:30
International Food Festival
Join us in the exhibit hall as you take a culinary trip around the world, sampling beers paired with foods from countries around the globe, near and far. Sponsored by

Thursday, April 2, 2015

7:30
Registration and Coffee

8:00
Chairperson's Remarks
Arvind Srivastava, Ph.D., Director, Formulation Development, Eli Lilly & Co.

Aggregation and Subvisible Particle Characterization and Control

8:15
Characterization of Aggregates in Blood-Derived Products and Their Recombinant Analogs
CDER and CBER recently collaborated to create a new Guidance for Industry document entitled "Analytical Procedures and Methods Validationfor Drugs and Biologics" which is published as a draft in February, 2014. This guidance was designed to update agency thinking on requirements for regulatory submission of analytical procedures and method validation data to support the documentation of the identity, strength, quality, purity, and potency of drug substances and drug products. The draft guidance supersedes the 2000 draft guidance for industry on Analytical Procedures and Methods Validation and, when finalized, will also replace the 1987 FDA guidance for industry on Submitting Samples and Analytical Data for Methods Validation. The agency also wanted to ensure this draft guidance would complement the International Conference on Harmonization (ICH) guidance documents, especially Q2 (R1) Validation of Analytical Procedures: Text and Methodology (Q2(R1)) for developing and validating analytical methods. In addition to assuring that guidance documents are current, the need for an updated guidance stems from the need to ensure analytical methodologies are consistent with changes to drug development and regulation seen with the concepts outlined in ICH Q8 (R2), Q9, Q10 and Q11.
Lokesh Bhattacharyya, Lab Chief, LACBRP/DBSQC, FDA/CBER/OCBQ (Invited)

8:45
Probing Structurally-Altered and Aggregated States of Therapeutically Relevant Proteins Using GroEL Coupled to Bio-Layer Interferometry
Nature employs chaperone proteins to detect and reverse large scale aggregation in vivo by interacting with partially folded protein species. Using this principle, we developed a GroEL chaperonin-Biolayer Interferometry biosensor to detect initial preaggregates in therapeutic protein formulations prior to large aggregate formation in vitro. We also determine if pre-aggregates detected by the GroEL-biosensor correlates with long term aggregation.
Mark Fisher Ph.D., Professor, Department of Biochemistry and Molecular Biology, Robert E. Hemenway Life Sciences Innovation Center, University of Kansas Medical Center

9:15
Monitoring and Control of Particulates in Protein Therapeutics
Particulates pose major concern in biological therapeutic development because of their potential to cause undesirable immunogenic reactions and other safety concerns. The particulates testing, monitoring, characterization and control strategy will be discussed in this presentation.
Arvind Srivastava, Ph.D., Director, Formulation Development, Eli Lilly & Co.

9:45
Networking Refreshment Break in Poster and Exhibit Hall

Aggregation and Subvisible Particle and Control (continued)

10:30
Analysis of Maytanisnoid ADCs by Imaged Capillary Isoelectric Focusing (icIEF)
Joyce Lin, MSc., Pr. Analytical Associate, Analytical and Pharmaceutical Sciences, ImmunoGen Inc.

11:00
Characterization of Potential Degradants in a 20 kDa PEGylating Raw Material by Orthogonal Analytical Techniques (RP-HPLC, APCI-MS and NMR)
Ying Luo, Process and Product Development, Amgen, Inc.

11:30
Regulatory Pathways for Licensure of Pandemic Influenza Vaccines: FDA Approach
David S. Cho, Ph.D., Senior Scientist for Emerging & Pandemic Threat Preparedness, CBER, U.S. FDA

Technology Workshop Presentation

12:05
Presentation TBA

The Secret Life of Protein A
Protein A chromatography is much more interesting and complex than its simple operation suggests. This presentation will reveal new data about conformational changes at the IgG-protein A interface, and describe newly discovered pathways by which eluted IgG becomes contaminated, by which aggregates are formed, by which IgG is lost at the protein A step — and how to prevent those problems.
Pete Gagnon, Bioprocessing Technology Institute, Singapore

12:35
Networking Luncheon in Poster and Exhibit Hall

1:40
Chairperson's Remarks
Jeffrey W. Hudgens, Ph.D., Research Chemist, NIST

Operational and Technology Considerations for Product and Process

Featured Presentation

1:45
Don't Go Into the Light! Analytical Characterization of a Photosensitive Therapeutic IgG1 Antibody After Exposure to Light
Galahad Deperalta, Scientist, Protein Analytical Chemistry, Genentech, a Member of the Roche Group, a member of the Roche Group

2:15
Strategies for Increasing Capacity and Capability of an Analytical Organizational Through Improved Technologies and Business Processes
Johnson Varghese, Senior Director, Head of Analytical Development, Shire HGT

2:45
Networking Refreshment Break and Last Chance for Poster and Exhibit Viewing

Methods and Strategies for Glycan Analysis

3:30
Comparison of Glycoproteins by Hydrogen-Deuterium Exchange-Electron Transfer Dissociation-Mass Spectrometry
Hydrogen-deuterium exchange mass spectrometry measures exchange rates of amide hydrogens along protein chains. Since hydrogen bonding associated with the higher order structure of glycoproteins can greatly slow amide hydrogen exchange, small changes in protein tertiary structure can change the H/D exchange profile. We demonstrate that changes in the glycan structure can also measurably change the HDX-MS patterns of glycoproteins.
Jeffrey W. Hudgens, Ph.D., Research Chemist, Institute for Bioscience and Biotechnology Research (CARB II/2204B), BioProcess Measurements Group, Biomolecular Measurement Division, National Institute of Standards and Technology (NIST)

4:00
Comprehensive N-Glycosylation Characterization of Biopharmaceuticals
This presentation will cover the state of the art of liquid phase N-glycan separation methods, mostly focusing on capillary electrophoresis and its combination with mass spectrometry for the analysis of biopharmaceuticals. Comprehensive N-glycosylation analysis of this recently emerging class of very successful new generation drugs will be discussed with the main goal to demonstrate structural and functional equivalence of innovator and biosimilar products.
Andras Guttman, Ph.D., Professor, Horvath Laboratory of Bioseparation Sciences, University of Debrecen, MMKK, Hungary

4:30
Glycan Analysis: An Updated Method for Existing Products
As knowledge about therapeutic recombinant proteins increase, minor glycan species have been identified as critical quality attributes. Newer technologies provide higher resolution and the ability to measure a greater number of glycan species, allowing improved product quality control. In this presentation, considerations for implementing newer analytical technologies for glycan measurement will be discussed.
Deanna Hurum, Ph.D., QC Scientist, Global Method Management and Technology, Genentech, a Member of the Roche Group

5:00
Close of BDP Week

BPI 2015
Charles River

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