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CE in the Biotechnology &
Pharmaceutical Industries:
18th Symposium on the Practical
Applications for the Analysis of
Proteins, Nucleotides and Small
Molecules
(CE Pharm 2016)
Symposium Co-Chairs:
Steffen Kiessig, F. Hoffmann – La Roche Ltd.
Henry Luo, Regeneron Pharmaceuticals, Inc.
September 25-28, 2016
The Westin San Diego
San Diego, CA
Organized by
2
Table of Contents
Welcome Letter .......................................................................................................... 3
CE Pharm Award ....................................................................................................... 4
Student Travel Grants ................................................................................................ 5
Program Partners, Exhibitors and Media Partners ..................................................... 6
Scientific Final Program Summary ............................................................................ 9
Session Abstracts ..................................................................................................... 17
Workshop I Description ........................................................................................... 44
Round Robin Table Discussions .............................................................................. 46
Technical Seminar Abstract ..................................................................................... 48
Poster Abstracts ....................................................................................................... 52
3
Welcome to CE Pharm 2016: CE in the Biotechnology and Pharmaceutical Industries: 18th
Symposium on the Practical Applications for the Analysis of Proteins, Nucleotides and Small
Molecules
We are pleased to welcome you to CE Pharm 2016, a symposium devoted to the practical concerns that
will strengthen the use of CE within the biotechnology and pharmaceutical industries. The goal of this
symposium is to provide a forum for the discussion of recent developments in the analysis and
characterization of protein therapeutics, nucleotides and small molecules by CE and related techniques.
The symposium will feature presentations from leading experts within industry and regulatory agencies
from around the world. Applications will highlight the use of CE in various areas of product development
including high-throughput screening, process development, product characterization, formulation studies,
validated lot release and stability testing. Attendees will have the opportunity to discuss the use of CE
with regulatory agencies. In addition, CE troubleshooting approaches will be presented and
instrumentation companies will show advances in CE instruments, sensitivity and reagents. The
symposium will allow for open discussions aimed at improving and increasing the use of CE for analysis
of proteins, small molecules, carbohydrates, metabolites, and other molecules, with a focus on validation
and qualification, new technology and QbD.
The success of this symposium will depend not only on the outstanding cast of experienced and
knowledgeable speakers and workshop leaders, but also on the interactions and open discussions that take
place among the attendees. We encourage you to participate whole-heartedly in the discussion sections
that have been designed to stimulate the exchange of ideas and information.
We would like to thank the speakers who are generously giving their time and resources and also you for
your attendance, which will make this endeavor a success.
We gratefully acknowledge the generosity of our exhibitors and program partners: 908 Devices,
Advanced Electrophoresis Solutions, Agilent Technologies, American Laboratory/Labcompare, American
Pharmaceutical Review, Amgen Inc., Analyst, Analytical Methods, The Analytical Scientist, Biogen,
BiOptic Inc., BioProcess International, ChemComm, Chemical Science, ChemSocRev, CMP Scientific,
Eli Lilly and Company, Genentech, a Member of the Roche Group, Genetic Engineering &
Biotechnology News, Green Chemistry, Integrative Biology, International Pharmaceutical Quality,
LCGC, MedChemComm, The Medicine Maker, Molecular BioSystems, The Pathologist, PerkinElmer,
Pfizer, Inc., Prince Technologies B.V., ProteinSimple, ProZyme, Regeneron Pharmaceuticals, Inc., Royal
Society of Chemistry, seperationsNOW.com, SCIEX, Technology Networks and Thermo Fisher
Scientific.
We are thankful for the expert assistance of CASSS and the audiovisual expertise of Michael Johnstone
from MJ Audio-Visual Productions. Their experience and guidance in the preparation of this symposium
have been invaluable.
THE SCIENTIFIC ORGANIZING COMMITTEE
Tim Blanc, Eli Lilly and Company
François de l'Escaille, ANALIS s.a./n.v.
Mei Han, Amgen Inc.
Göran Hübner, Boehringer Ingelheim Pharma
GmbH & Co. KG
Steffen Kiessig, F. Hoffmann-La Roche Ltd. (Co-chair)
Nathan Lacher, Pfizer, Inc.
C. Mark Lies, SCIEX
Henry Luo, Regeneron Pharmaceuticals (Co-chair)
David A. Michels, Genentech, a Member of the Roche
Group
SungAe Suhr Park, Amgen Inc.
Richard Rustandi, Merck & Co., Inc.
Oscar Salas-Solano, Seattle Genetics, Inc.
Cari Sänger - van de Griend, Kantisto BV
Zoran Sosic, Biogen
Hermann Wätzig, Technical University Braunschweig
Joel Welch, CDER, FDA
4
CE Pharm Award History and Qualifications
Objective: Recognize and award an individual for sustained and significant contribution to the practical
application of CE to the analysis of biotechnology and pharmaceutical products.
Qualification for Award:
a. Advocate for CE from biotechnology and pharmaceutical industry b. Technical advancement or considered as a leader in developing or implementing
various CE applications, such as:
New CE Application for R&D
CE Method Qualification
CE Method Validation
CE Method Transfer
c. Technical reputation, in terms of number of presentations, publications, and
patents
d. Dedication to CE Pharm meeting as speaker, tutor, poster presenter or committee
member
e. Mentor, advisor and advocate of industrial-based CE practitioners in other
industrial applications such as food chemistry, forensics and clinical.
Past Recipients of the "CE Pharm Award" include:
2006 - Norberto Guzman – Johnson & Johnson
2007 - Kevin Altria – GlaxoSmithKline
2008 - Anthony Chen and Wassim Nashabeh – Genentech, Inc.
2009 - Stacey Ma – Genentech, Inc.
2010 - SungAe Suhr Park – Amgen Inc.
2011 - Oscar Salas-Solano – Seattle Genetics, Inc.
2012 - Franka Kálmán – University of Applied Sciences Western Switzerland
2013 - András Guttman – Northeastern University
2014 - Michel Girard – Health Canada
2015 - Cari Sänger - van de Griend – Kantisto BV
2016 - Winner will be announced Wednesday at 10:10 AM.
Do you think we are missing someone influential? Add your suggestion to the list.
Suggestions for next year’s award can be submitted with your post-meeting evaluation.
5
CASSS CE Pharm Student Travel Grants
CASSS is pleased to provide a limited number of student travel grants for students who present
applicable posters at CE Pharm 2016. PhD students or post-doctoral fellows conducting research
in academia or industry throughout the world are eligible.
Why you should apply:
This symposium gives insight into the current topics and issues under discussion within the
pharmaceutical and biotech industry and, as such, gives attendees the opportunity to bridge
between industry, academia and regulatory agencies. The presentations and workshops will be
devoted to practical concerns that strengthen the use of CE within the biotechnology and
pharmaceutical industries. Applications will highlight uses of CE in various areas of product
development, including high-throughput screening, formulation studies, process development,
product characterization and validated lot release and stability testing. As a participant, you will
have an excellent opportunity to meet, network and participate in exchanging knowledge for
mutual education with other CE practitioners.
Requirements are:
- Present a poster on a CE topic
- Proof of studentship/post-doc status
- Recommendation from the supervisor/advisor
CASSS has awarded student travel grants to the following individuals:
Simultaneous Determination of Protein Affinity and Heterogeneity by Capillary
Electrophoresis–Mass Spectrometry
Rob Haselberg, Vrije Universiteit Amsterdam, Netherlands
Affinity Capillary Electrophoretic and Computational Methods for Binding Studies of P-
selectin with Heparinoids
Mona Mozafari, TU Braunschweig, Germany
Capillary Zone Electrophoresis-electrospray Ionization-mass spectrometry for Xenopus
Laevis Metabolomic Analysis
Nicole Schiavone, University of Notre Dame, USA
Coupling Capillary Zone Electrophoresis to a Q Exactive HF Mass Spectrometer for Top-
down Proteomics: 580 Proteoform Identifications from Yeast
Yimeng Zhao, University of Notre Dame, USA
6
The Scientific Organizing Committee gratefully acknowledges the following
strategic partners for their generous support of this Symposium:
Strategic Program Partners
Diamond
Genentech, a Member of the Roche Group
Platinum
Biogen
Gold
Pfizer, Inc.
Diamond Program Partner
ProteinSimple
Platinum Program Partner
SCIEX
Gold Program Partners
Agilent Technologies
PerkinElmer
Silver Program Partners
Amgen Inc.
Eli Lilly and Company
7
Welcome Reception
Regeneron Pharmaceuticals
Exhibitors
908 Devices Inc.
Advanced Electrophoresis Solutions
Agilent Technologies
BiOptic Inc.
CMP Scientific
PerkinElmer
ProteinSimple
Prince Technologies B.V.
ProZyme
SCIEX
Thermo Fisher Scientific
8
The Scientific Organizing Committee gratefully acknowledges the following
media for their promotional consideration of CE Pharm 2016:
Media Program Partners
American Laboratory/labcompare
American Pharmaceutical Review
Analyst
Analytical Methods
The Analytical Scientist
BioProcess International
ChemComm
Chemical Science
Chem Soc Rev
Genetic Engineering & Biotechnology News
Green Chemistry
Integrative Biology
International Pharmaceutical Quality
LCGC
MedChemComm
The Medicine Maker
Molecular BioSystems
The Pathologist
Royal Society of Chemistry
separationsNOW.com
Technology Networks
9
CE Pharm 2016
Scientific Final Program Summary
Sunday, September 25, 2016
08:30 – 09:00 Breakfast (for course attendees ONLY) in the Pearl Room
08:30 – 13:00 Registration (for course attendees ONLY) in the 3rd Floor Foyer
09:00 – 17:00
Short Course in Pearl Room
Applications of Capillary Electrophoresis to the Analysis of Protein Therapeutics
Short Course Facilitators:
David A. Michels, Genentech, a Member of the Roche Group, South San Francisco, CA USA
and Cari Sänger - van de Griend, Kantisto BV, Baarn, The Netherlands
10:30 – 11:00 Break in the Pearl Room
12:30 – 13:30 Hosted Lunch (for course attendees ONLY) on the Pool Deck
15:00 – 15:30 Break in the Pearl Room
16:30 – 17:30 Registration CE Pharm 2016 in the Ballroom Foyer
17:00 – 19:00 Welcome Reception at the Ivory Room and Pool Deck
10
Monday, September 26, 2016
07:30 – 18:00 Registration in the Ballroom Foyer
07:30 – 08:30 Breakfast in Emerald Ballroom
08:30 – 08:45 Welcome and Introductory Comments in Crystal Ballroom
Henry Luo, Regeneron Pharmaceuticals, Inc., Tarrytown, NY USA
Keynote I Session in Crystal Ballroom
Session Chair: Henry Luo, Regeneron Pharmaceuticals, Inc., Tarrytown, NY USA
08:45 – 09:30 Control Strategy as a Framework for Integrated Analytical and
Process Development of Biotherapeutics and Vaccines
Margaret Ruesch, Pfizer, Inc., Chesterfield, MO USA
09:30 – 09:45 Discussion
09:45 – 10:15 Break – Visit the Exhibits and Posters in Emerald Ballroom
Protein Analysis and Emerging Therapeutics in Crystal Ballroom
Session Chairs: Richard Rustandi, Merck & Co., Inc., West Point, PA USA
and Hermann Wätzig, Technical University Braunschweig, Braunschweig, Germany
10:15 – 10:40 Purity Analysis of Bispecific Antibody by Affinity Capillary
Electrophoresis
Kathir Muthusamy, Regeneron Pharmaceuticals, Inc., Tarrytown, NY USA
10:40 – 11:05 High-throughput Analysis of Antigen-specific, Vaccine-induced
Antibody Glycosylation using multi-capillary CE Todd Suscovich, Ragon Institute of MGH, MIT, and Harvard University,
Cambridge, MA USA
11:05 – 11:30 Novel CZE Method for the Quantification of Intact Virus Particles in
Complex Matrices – Quality by Design Method Development and
Implementation in a GMP Environment
Ewoud van Tricht, Janssen Infectious Diseases and Vaccines, Leiden,
Netherlands
11:30 – 11:55 Evaluation of Capillary Electrophoresis as a Potential Alternative
Method to Monitoring Fragment Levels in Antibody-drug Conjugates
Nomalie Jaya, Seattle Genetics, Inc., Bothell, WA USA
11:55 – 12:10 Discussion
11
Monday, September 26, 2016 continued
12:10 – 12:25 Lunch for Technical Seminar Attendees – Please take lunch and return
to Crystal Ballroom for the “Lunch and Learn”
12:25 – 13:25 Technical Seminar/Lunch and Learn
Run Your Assays Faster: Getting the Quantitative Answers You Need For N-Glycan, CE-
SDS, and CZE Applications Without the Wait
Matt Salem, SCIEX, Brea, CA USA
Sponsored by SCIEX Crystal Ballroom
13:25 – 13:40 Mini Break – Visit the Exhibits and Posters in Emerald Ballroom
QbD and DoE for CE Session in Crystal Ballroom
Session Chairs: Tim Blanc, Eli Lilly and Company, Branchburg, NJ USA
and Göran Hübner, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany
13:40 – 14:05 Development and Validation of a Capillary Gel Electrophoresis Purity
Assay for a Therapeutic Protein using the Quality by Design
Approach: A Step Further in Analytical Lifecycle Management
Jérémie Cuisenaire, UCB Pharma SA, Braine-l'Alleud, Belgium
14:05 – 14:30 Optimization of Capillary Zone Electrophoresis for Charge
Heterogeneity Testing of Biopharmaceuticals using Enhanced Method
Development Principles
Bernd Moritz, F. Hoffmann - La Roche Ltd., Basel, Switzerland
14:30 – 14:55 Automated DOE for iCIEF Method Development
Peter Bryngelson, Biogen, Cambridge, MA USA
14:55 – 15:20 Quality by Design in the Development of Capillary Electrophoresis
Methods for Recombinant Proteins
Agatha Feltus, Elanco Animal Health, Greenfield, IN USA
15:20 – 15:35 Discussion
15:35 – 16:05 Break – Visit the Exhibits and Posters in Emerald Ballroom
16:05 – 17:00
CE Pharm Partner Showcase in Crystal Ballroom
Facilitator: SungAe Suhr Park, Amgen Inc., Thousand Oaks CA USA
12
Monday, September 26, 2016 continued
17:00 – 18:00
Workshop I: Troubleshooting in Crystal Ballroom
Workshop Facilitators: Tim Blanc, Eli Lilly and Company, Branchburg, NJ USA
and Cari Sänger - van de Griend, Kantisto BV, Baarn, Netherlands
18:00 – 19:00 Exhibition Reception – Visit the Exhibitors in Emerald Ballroom
13
Tuesday, September 27, 2016
08:00 – 18:00 Registration in the Ballroom Foyer
07:30 – 08:30 Breakfast in Emerald Ballroom
07:30 – 08:30 1st Time Attendee Breakfast in Emerald Ballroom
Keynote II Session in Crystal Ballroom
Session Chair: SungAe Suhr Park, Amgen Inc., Thousand Oaks, CA USA
08:30 – 09:15 High-Throughput Protein, Protein-Protein Interaction, and
Metabolite Assays using CE
Robert Kennedy, University of Michigan, Ann Arbor, MI USA
09:15 – 09:30 Discussion
09:30 – 10:00 Break – Visit the Exhibits and Posters in Emerald Ballroom
Deep Dive into CE Session in Crystal Ballroom
Session Chairs: David Michels, Genentech, a Member of the Roche Group, South San Francisco,
CA USA and Cari Sänger - van de Griend, Kantisto B.V., Baarn, Netherlands
10:00 – 10:25 cIEF – Resolution versus pI Accuracy
Gordon Freckleton, Eli Lilly and Company, Branchburg, NJ USA
10:25 – 10:50 Studying Aggregation of Proteins Using Capillary Electrophoresis
Akram Khodabandehloo, UBC, Vancouver, BC Canada
10:50 – 11:15 Assessing Critical Quality Attributes of Therapeutic Proteins by Size-
Based Electrophoretic Separation Methods: A Deep Dive into CE!
Thomas Niedringhaus, Genentech, a Member of the Roche Group, South
San Francisco, CA USA
11:15 – 11:30 Discussion
11:30 – 11:45 Lunch for Technical Seminar Attendees – Please take lunch and return
to Crystal Ballroom for the “Lunch and Learn”
11:45 – 12:45 Technical Seminar/Lunch and Learn
Meet Maurice, One-stop cIEF and CE-SDS for Y our Biologics
Xin Jiang, ProteinSimple, San Jose, CA USA
Jiaqi Wu, ProteinSimple, Toronto, ON Canada
Sponsored by ProteinSimple Crystal Ballroom
14
Tuesday, September 27, 2016 continued
12:45 – 13:00 Mini Break – Visit the Exhibits and Posters in Emerald Ballroom
Novel Technologies Session in Crystal Ballroom
Session Chairs: Mei Han, Amgen Inc., South San Francisco, CA USA
and Nathan Lacher, Pfizer, Inc., Chesterfield, MO USA
13:00 – 13:25 Mass Spectrometric Characterization of Impurities of an Antibody
Separated by SDS-capillary Sieving Electrophoresis using CSE-CZE-
MS
Christian Neusüß, Aalen University, Aalen, Germany
13:25 – 13:50 cIEF Cross Platform Comparability Study of a Monoclonal Antibody
John Barr, Amgen Inc., South San Francisco, CA USA
13:50 – 14:15 Current Developments in CE/MS Analysis of Proteins
Michael Knierman, Eli Lilly and Company, Indianapolis, IN USA
14:15 – 14:40 Knock! Knock! Who’s there? “Maurice” Who?
SungAe Suhr Park, Amgen Inc., Thousand Oaks, CA USA
14:40 – 14:55 Discussion
14:55 – 16:50 Poster Session - Visit the Exhibits and Posters in Emerald Ballroom
16:50 – 17:50
Round Robin Table Discussion in Crystal and Emerald Ballroom
Roundtable Facilitator: François de l’Escaille, ANALIS s.a./n.v., Suarlee, Belgium
15
Wednesday, September 28, 2016
08:00 – 15:00 Registration in the Ballroom Foyer
07:30 – 08:30 Breakfast in the Diamond Room Foyer
Regulatory Session in Diamond Room
Session Chairs: Oscar Salas-Solano, Seattle Genetics, Inc., Bothell, WA USA
and Joel Welch, CDER, FDA, Silver Spring, MD USA
08:30 – 09:05 Now You CE Me, Now You Don’t: The Evolving Role of CE in the
Control Strategy Sarah Kennett, CDER, FDA, Silver Spring, MD USA
09:05 – 09:30 Application of Microchip Capillary Electrophoresis in the Quality
Control Laboratory
Jeffery Schneiderheinze, Regeneron Pharmaceuticals, Inc., Tarrytown, NY
USA
09:30 – 09:55 CE Applications in a QC Environment
Jian Zhang, Genentech, a Member of the Roche Group, South San
Francisco, CA USA
09:55 – 10:10 Discussion
10:10 – 10:20 Presentation of the CE Pharm Award
10:20 – 10:45 Break in the Diamond Room Foyer
Method Lifecycle Management Session in Diamond Room
Session Chairs: C. Mark Lies, SCIEX, Brea, CA USA
and Zoran Sosic, Biogen, Cambridge, MA USA
10:45 – 11:10 The NISTmAb: A Comprehensively Characterized Reference
Material to Support Biopharmaceutical Analytical Technology
Development
Abigail Turner, Institute for Bioscience and Biotechnology Research
(IBBR), Rockville, MD USA
11:10 – 11:35 CE Analysis in GMP Environment – What is that Noise?
Jiu-Li Song, Amgen Inc., Thousand Oaks, CA USA
11:35 – 12:00 Development on an icIEF Method for a Poorly Soluble ADC Derived
from an IgG2 mAb Conjugated with a Charged Drug-linker
Adam Fung, Agensys, Inc., Santa Monica, CA USA
16
Wednesday, September 28, 2016
12:00 – 12:15 Discussion
12:15 – 12:30 Lunch for Technical Seminar Attendees – Please take lunch and return
to Diamond Room for the “Lunch and Learn”
12:30 – 13:10 Technical Seminar/Lunch and Learn
High Throughput and High Resolution N-glycan Analysis Using Multicapillary CE and
Novel Fluorescent Dyes
Shaheer Khan, Thermo Fisher Scientific, South San Francisco, CA USA
Sponsored by Thermo Fisher Scientific Diamond Room
13:10 – 13:20 Mini Break – Please proceed to Crystal Ballroom
CE-MS Session in Crystal Ballroom
Session Chairs: Steven Cohen, Northwestern University, Boston, MA USA
and Steffen Kiessig, F. Hoffmann - La Roche Ltd., Basel, Switzeraland
13:20 – 14:00 Microfluidic Capillary Electrophoresis-Electrospray Devices for
Analysis of Biopharmaceutical Materials
J. Michael Ramsey, University of North Carolina, Chapel Hill, Chapel
Hill, NC USA
14:00 – 14:30 Cutting-Edge Mass Spectrometry for mAbs & Related Product
Structural Characterization
Elsa Wagner-Rousset, Centre d'Immunologie Pierre Fabre, Saint Julien
en Genevois, France
14:30 – 15:00 CE-MS Analysis of A Single Dose Pharmacokinetic Study of Two Fc-
Fusion Protein Constructs
Mei Han, Amgen Inc., South San Francisco, CA USA
15:00 – 15:15 Closing Comments in Crystal Ballroom
Steffen Kiessig, F. Hoffmann - La Roche Ltd., Basel, Switzerland
17
Oral Abstracts
Control Strategy as a Framework for Integrated Analytical and Process Development of
Biotherapeutics and Vaccines
Margaret Ruesch
Pfizer, Inc., Chesterfield, MO USA
Analytical organizations play a central role in developing biotherapeutic and vaccine candidate
molecules and manufacturing processes for the patients we serve. A core mission of a
Biotherapeutics Pharmaceutical Sciences Analytical organization is the development of Quality
Attribute understanding as well as the stewardship of this knowledge across cross-functional
teams. Quality Attribute knowledge is the foundation for developing phase-appropriate process
understanding, and underpins the establishment of Control Strategies per ICH Q10.
Efforts to simplify Quality Attribute and Control Strategy approaches and tools are critical to
ensure we are focusing our scientific efforts on the most important aspects of a product
throughout all stages of development. In addition, these simplified risk-based approaches offer a
framework for ensuring the successful development of complex modalities and accelerated
programs. In early phases, the use of platform analytical methods and strategies, together with
the targeted application of enhanced analytical characterization, forms the foundation for early
process and product development. By later phases, more extensive product and process
understanding needs to be translated into an understandable, robust, and defendable Control
Strategy. Case studies where platform approaches and enhanced analytical characterization have
been applied to develop phase-appropriate Control Strategies will be discussed. Looking
forward, technical and organizational efforts to move Quality Attribute understanding closer to
process, as opposed to traditional reliance on retrospective testing, are particularly critical for
further integration of analytical and process development and for improvements in efficiency,
speed and quality.
NOTES:
18
Purity Analysis of Bispecific Antibody by Affinity Capillary Electrophoresis
Kathir Muthusamy, Henry Luo, Erica Pyles
Regeneron Pharmaceuticals, Inc., Tarrytown, NY USA
Despite wide therapeutic applications for bispecific antibodies (bsAbs), challenges associated
with manufacture and purity analysis prevail. Co-expression of two heavy chains with a common
light chain minimizes the number of monospecific antibodies (from 8 to 2) that are subsequently
removed during purification. Conventional purity analysis methods may be inadequate due to
similar physiochemical properties of bispecific and monospecific antibodies that may be present
following expression. To address this challenge, a robust and powerful Capillary Zone
Electrophoresis (CE) method combined with affinity CE has been developed.
NOTES:
19
High-throughput Analysis of Antigen-specific, Vaccine-induced Antibody Glycosylation
using Multi-capillary CE
Todd Suscovich
Ragon Institute of MGH, MIT, and Harvard University, Cambridge, MA USA
Antibody effector functions, such as antibody-dependent cellular cytotoxicity, antibody-
dependent complement deposition, and antibody-dependent phagocytosis, play a critical role in
immunity against multiple pathogens, particularly in the absence of neutralizing activity. One of
the principal methods by which antibody functionality is regulated in the body is via changes to a
single N-linked glycan located in the CH2 domain of the IgG Fc. Over 30 different glycan
structures can be present at this site, and the removal of specific sugar moieties can have a
dramatic effects on the functions associated with a given antibody. For example, the removal of
fucose results in dramatically increased FcγR3a-dependent antibody-dependent cellular
cytotoxicity, whereas the removal of galactose can result in in enhanced complement binding and
deposition. Studies of IgG glycosylation in vivo have been historically limited by the low-
throughput nature of existing analytical techniques, which generally require prohibitively
expensive instrumentation and large quantities of sample, thus limiting the scope of research into
both natural regulation of IgG glycosylation as well as vaccine-induced changes in IgG
glycosylation. Traditional approaches to analyze IgG glycosylation have relied primarily on high
performance liquid chromatography or mass spectrometry, both of which require relatively large
quantities of material/antibody for accurate analysis as well as significant time and expertise to
acquire and analyze data. As studies of IgG glycosylation begin to focus on in vivo glycan
modifications, both in human populations and in animal models, the sample volume available for
analysis often becomes limiting while the number of samples that need to be analyzed increases
exponentially. Furthermore, because the vaccine specific-antibodies account for only a few
percent of all circulating antibodies, the analytical methods must be able to evaluate antibody
glycosylation on remarkably small amounts of material. Capillary electrophoresis offers a unique
high-throughput, quantitative analytical tool for the analysis of low-abundance antibody
glycosylation profiles. Coupled recent advances in glycan release, labelling, and purification, the
use of common DNA sequencing equipment to perform glycan structure analysis by capillary
electrophoresis is an excellent alternative to the established methods, with advantages in
simplicity, throughput, structural resolution, and sensitivity.
NOTES:
20
Novel CZE Method for the Quantification of Intact Virus Particles in Complex Matrices –
Quality by Design Method Development and Implementation in a GMP Environment
Ewoud van Tricht1, Lars Geurink1, Cari Sänger – van de Griend2
1Janssen Infectious Diseases and Vaccines, Leiden, Netherlands, 2Kantisto BV, Baarn,
Netherlands
Quantification of the intact adenovirus particle concentration is needed to support manufacturing
process development. The current methods are either not sufficiently accurate due to carry-over
and recovery issues, or require very long analysis times in order to reach adequate analytical
precision. Therefore capillary electrophoresis (CE) was evaluated for fast, accurate and precise
quantification of adenovirus particles.
An analytical quality by design (AQbD) method development approach was embraced. With CE,
the intact adenovirus particles were separated from sample matrix components such as cell
debris, residual cell DNA, proteins, and/or salts. The background electrolyte (BGE) composition,
analysis time, and sample pretreatment were optimized using a full factorial design of
experiments. BGE additives, capillary temperature, the use of a coated capillary and capillary
conditioning were investigated and proved vital to reduce virus adsorption, particulate matter and
carry-over, and to allow long series of measurements.
The method was validated for the quantification of adenoviruses in the range of 80 – 250 pmol/l
adenovirus particles (0.5 x1011 to 1.5 x1011 particles/ml) for representative samples from
downstream and upstream processing. The intact adenovirus particle concentrations obtained by
CE were overall equivalent to those obtained by quantitative polymerase chain reaction (qPCR),
although the CE method showed better precision (RSD < 6%) than anion-exchange HPLC and
qPCR (10 – 25% RSD). Additionally, analysis times were much shorter with CE than with
qPCR, allowing analysis of 30 samples in less than 4 hours. CE proved highly useful for process
development support and is implemented for in-process control testing for adenovirus vaccine
manufacturing.
NOTES:
21
Evaluation of Capillary Electrophoresis as a Potential Alternative Method for Monitoring
Fragment Levels in Antibody-Drug Conjugates
Nomalie Jaya
Seattle Genetics, Inc., Bothell, WA USA
Low Molecular Weight (LMW) fragments are important product quality attributes routinely
monitored in monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs). Size-
Exclusion Chromatography (SEC) has been the method of choice for evaluating fragments.
However, SEC does not always provide adequate resolution of a common antibody fragment
generated via hinge cleavage resulting in a FcFab (~100Kda) and Fab (~50Kda) species which
may contribute to a decrease in target binding resulting in reduced product efficacy.
Capillary Electrophoresis-Sodium Dodecyl Sulfate (CE-SDS) assays, while relying on different
separation principals from SEC, is an orthogonal method for monitoring fragments. Additionally,
CE separation can provide improved fragment resolution leading to a more robust and
reproducible quantitation. By comparing the fragmentation patterns between SEC and CE-SDS
we have demonstrated that both methods can accurately capture fragmentation trends resulting
from antibody hinge cleavage. We propose to leverage this information to evaluate whether CE-
SDS could be used as an alternative method for monitoring fragments levels in ADCs.
NOTES:
22
Development and Validation of a Capillary Gel Electrophoresis Purity Assay for a
Therapeutic Protein using the Quality by Design Approach: A Step Further in Analytical
Lifecycle Management
Jérémie Cuisenaire, Marc Jacquemin
UCB Pharma SA, Braine-L’Alleud, Belgium
Implementing analytical Quality by Design principles to Biologics analysis is a challenge due to
protein complexity (structure, micro-heterogeneity…). However, the benefits of applying the
QbD principles to analytical methods are clearly seen in terms of enhancing the understanding,
the robustness and the control of the method throughout its lifecycle.
The analytical QbD process consists of various steps and starts with the definition of an
Analytical Target Profile (ATP) based on molecular characteristics. Following the technology
selection, a risk assessment using different tools (Fishbone diagram, FMEA...) is performed.
Design of Experiments (DoE) studies are at the heart of this methodology for identifying and
understanding method variability and establishing method parameters which provide desirable
method performance. After determining the final analytical conditions, the validation is carried
out using the accuracy profile strategy.
This presentation reports the different steps of the QbD methodology that were applied to the
development and validation of a capillary gel electrophoresis purity assay in non-reducing
conditions for a therapeutic protein.
NOTES:
23
Optimization of Capillary Zone Electrophoresis for Charge Heterogeneity Testing of
Biopharmaceuticals using Enhanced Method Development Principles
Bernd Moritz1, Valentina Locatelli2, Barbara Entler2, Michele Niess1, Rolf Ketterer1, Andrea
Heyne1, Steffen Kiessig1
1F. Hoffmann - La Roche Ltd., Basel, Switzerland, 2IMC Fachhochschule Krems, Krems an der
Donau, Austria
Capillary zone electrophoresis is a powerful technique for charge heterogeneity testing of
biopharmaceuticals [1]. It is based on the differences between the ratios of net charge and
hydrodynamic radius. In an extensive intercompany study it was recently shown that capillary
zone electrophoresis is very robust and can be easily implemented in labs that didn’t perform it
before [2].
The distribution of different charge species strongly depends on the individual characteristics of
the examined protein that sometimes resulted in suboptimal resolution. Enhanced method
development principles where applied to investigate possibilities for further method
optimization. For this purpose the different method parameters and ingredients (factors) were
investigated in several DOE studies. That resulted in some factors that are most important for
improving CZE separation. For this set of factors DOE models were generated and optimized in
several ways for different sets of responses out of 14 target criteria like resolution, peak width
and number of peaks. In spite of product specific DOE optimization it was found that the
resulting combination of factors did also result in significant improvement for 17 of 19 different
antibodies or product formats. This clearly demonstrates some generic applicability of this
method. However, adaptation to individual molecular properties is sometimes still required in
order to achieve best results. The set screws that were found in this study are well suited for this
specific optimization and are expected to significantly reduce effort for this final step.
[1] Y. He et al., J. Sep. Sci. 2011, 34, 1-8
[2] B. Moritz et al., J. Chrom. B, 2015, 983-984, 101-110
NOTES:
24
Automated DOE for iCIEF Method Development
Peter Bryngelson, Richard Smart
Biogen, Cambridge, MA USA
Thousands of protein charge variant samples may be analyzed during the progression of a bio-
therapeutic throughout the various stages of discovery and clinical development. Imaged
capillary isoelectric focusing (iCIEF) instrumentation has the capability of analyzing up to 96
samples in approximately 24 hours, however sample preparation on that scale can be
cumbersome. In an effort to efficiently develop and qualify robust platform methods; liquid
handling automation and Design of Experiments (DoE) have been combined to develop
predictive models to accurately describe the interaction of relevant sample preparation factors.
Method development can be completed in as few as two experiments, with full ICH Q2 (R1)
qualification in as little as five experiments. The flexible automated approach also provides the
ability to analyze a wide variety of samples in a single run, bringing efficiencies to routine
testing as well as method development. Here we present the automated development of iCIEF
sample preparation conditions for a monoclonal antibody.
NOTES:
25
Quality by Design in the Development of Capillary Electrophoresis Methods for
Recombinant Proteins
Agatha Feltus
Elanco Animal Health, Greenfield, IN USA
Quality by design (QbD) in analytical development can be a powerful tool in the development of
robust methods. The QbD process is a systematic approach that includes: identifying critical
quality attributes (CQAs) to be measured, defining the performance requirements of the method
intended to measure them, assessing the risks of various method parameters, defining the
method’s design space and control space, and on-going monitoring for continuous improvement.
In this talk, we will walk through the process and apply the QbD approach to an example
capillary electrophoresis method by generating an Analytical Target Profile, performing a risk
assessment, and developing potential DOEs to optimize the method. Finally, we will consider
long-term performance evaluation in terms of appropriate control charting and system suitability.
NOTES:
26
High-Throughput Protein, Protein-Protein Interaction, and Metabolite Assays using CE
Robert Kennedy
University of Michigan, Ann Arbor, MI USA
A critical advantage of capillary and microchip electrophoresis (CE and MCE) is the potential
for high-speed separations. With high voltages over short distances (e.g., 1-3 kV/cm) it is
possible to obtain efficient separations in seconds. This speed can be used to improve throughput
and to capture non-covalent complexes. These advantages apply to both proteins and small
metabolites. In this talk we provide an overview of fast CE and MCE as well as the use of such
speed in new methods for protein analysis, protein-protein interaction (PPI) assay, high
throughput screening, and sensing. The most common protein analysis method is a western blot.
Despite its great popularity little work has been done on miniaturizing, automating, or improving
the speed of this method. We have interfaced MCE to a protein capture membrane to provide a
western blot that can be completed at much higher throughput than coventional westerns.
Another substantial advantage is the capability of multiplexing to determine multiple proteins
from single samples. In another direction, we have used rapid separations to detect PPI by
separating bound and free proteins. More recently we have developed protein-cross-linking CE
(PXCE) which allows a much wider variety of interacting proteins to be probed quantitatively by
CE. These methods may be used for screeing interacting proteins and for testing aggregation of
proteins. Although MCE allows fast separations, throughput can be limited by the speed of
sample introduction to a chip. We have used segmented flow, were samples are stored as
nanoliter droplets compartmentalized in an immiscible carrier fluid, to introduce samples rapidly
into a MCE chip. Sample analysis rates as high as 1/s for over 1000 samples can be achieved.
This technology is used for drug discovery screening and to analyze samples collected in vivo
for high temporal resolution "sensing".
NOTES:
27
cIEF – Resolution versus pI Accuracy
Gordon Freckleton, Tara Enda, Dana Lazich, Tim Blanc
Eli Lilly and Company, Branchburg, NJ USA
Capillary isoelectric focusing (CIEF) offers tremendous advantages over traditional IEF both in
terms of the ability to resolve individual charge isoforms and the ability to assign precise
isoelectric points to them. However, it is still limited by the quality of the carrier ampholytes
used to create the underlying gradient. We will look at the quality of ampholyte gradients
through the context of the Righetti lab’s exhaustive examination of commercially-available
narrow-range ampholytes. We will also look at the practical evaluation of medium- and broad-
range ampholytes in a CIEF context with an eye towards balancing resolution and isoelectric
point accuracy, and an approach to more accurately estimate isoform isoelectric points.
NOTES:
28
Studying Aggregation of Proteins Using Capillary Electrophoresis
Akram Khodabandehloo, David D.Y. Chen
UBC, Vancouver, BC Canada
The protein aggregation is an inevitable phenomenon in protein formulations which causes
inconsistency in the final product and also affect the efficacy and immunogenicity of protein-
based drugs. To confirm the safety of these products, they must be approved through regulatory
agencies before entering the market. Sizing techniques are good candidates for monitoring of the
protein aggregates since size is the main change in the aggregation of proteins.
Taylor Dispersion Analysis (TDA) is a recently developed method for estimating a wide range of
molecular and particle sizes, but it is not the best choice when dealing with the mixtures. To
overcome this limitation, we have combined TDA with the separation power of capillary
electrophoresis (CE) and used it to study a mixture of proteins and their aggregates. In this work,
the aggregates for BSA and IgG are made under heat stress and the size and charge are estimated
for monomers and the aggregates. The principle of method and the latest results will be
discussed.
NOTES:
29
Assessing Critical Quality Attributes of Therapeutic Proteins by Size-Based
Electrophoretic Separation Methods: A Deep Dive into CE!
Thomas Niedringhaus
Genentech, a Member of the Roche Group, South San Francisco, CA USA
For therapeutic proteins in late-stage development, understanding the impact of various process
parameters on critical quality attributes (CQAs) is a desired outcome to the quality-by-design
(QbD) manufacturing of therapeutic proteins prior to commercialization. QbD not only ensures
quality to patients, but it also leads to better understanding of the process which in turn
minimizes the probability of failed production lots. Throughout process characterization and
process validation studies, and for large-scale clinical campaigns, sensitive size-based
electrophoretic methods are employed to assess and monitor CQAs like product-related low-
molecular weight species (LMWS) and host-cell protein (HCP) impurities. This talk will take a
deep dive into the capabilities of reduced and non-reduced CE-SDS methods in understanding
the type of LMWS that are monitored for a given monoclonal antibody product, and their ability
to detect HCPs. In addition, the idea of an appropriate quality control system that reflects the
risks and the knowledge gained through QbD will be discussed.
NOTES:
30
Mass Spectrometric Characterization of Impurities of an Antibody Separated by SDS-
capillary Sieving Electrophoresis using CSE-CZE-MS
Christian Neusüß
Aalen University, Aalen, Germany
Capillary Electrophoresis is a key technology for the separation of variants and impurities of
therapeutic proteins. However, identification by e.g. mass spectrometry is difficult since on one
hand upscaling and fraction collection is difficult and on the other hand most methods require the
use of non-volatile and ESI-interfering constituents which prevent an on-line coupling to mass
spectrometry.
Recently we introduced the concept of two-dimensional CE applying a mechanical valve in order
to couple the first (non-MS-compatible) dimension in a heart-cut approach to CZE-MS. The
second dimension is used to separate the analytes of interest from all interfering constituents of
the first dimension using a MS-compatible BGE [1]. In this way it should be possible to couple
any kind of capillary-based electromigration technique to mass spectrometry. So far, this has
been demonstrated for CZE using phosphate buffer for peptide separation [1], CZE using tricine
buffer for the separation of degradation products of acetylsalicylic acid and ascorbic acid [2] as
well as capillary isoelectric focusing for protein separation [3]. Furthermore, the detailed mass
characterization of charge variants of intact antibodies separated in an ε-aminocaproic acid
(EACA) based BGE is possible.
In this presentation, a focus is set on the coupling of SDS-capillary sieving electrophoresis to
CZE-MS (CSE-CZE-MS). In this case, the SDS-containing protein from the first dimension has
to be in-line purified in the second dimension. In order to remove both SDS, non-volatile buffer
ions and the pseudo-gel matrix a method based on the co-injection of complexing agents and
organic solvents has been developed. This set-up is incorporated in the second dimension with
the valve containing a 20nl loop transferring the sample from the first to the second dimension.
In this way the CSE-CZE-MS approach allows for the first time the mass characterization of
impurities of antibodies separated by SDS-CSE.
[1] Kohl, Montealegre, Neusüß Electrophoresis (2016) Jan 22. doi: 10.1002/elps.201500579
[2] Neuberger, Jooß, Ressel, and Neusüß, Anal. Bioanal. Chem. (2016), DOI 10.1007/s00216-
016-9734-2
[3] Hühner, Neusüß, Anal. Bioanal. Chem. (2016), 408, 15, 4055–4061.
NOTES:
31
cIEF Cross Platform Comparability Study of a Monoclonal Antibody
John Barr
Amgen Inc., South San Francisco, CA USA
Abstract not available at time of print.
NOTES:
32
Current Developments in CE/MS Analysis of Intact Proteins
Michael Knierman, Megan Lannan
Eli Lilly and Company, Indianapolis, IN USA
CE/MS is gaining popularity mainly driven in improvement in mass spectrometry’s sensitivity.
My talk will cover the implementation of a nanosheath CE/MS interface. The interface was
placed on an Agilent 6550 and a Thermo orbitrap velos pro mass spectrometer using a
commercial Agilent CE system. The primary aim was to develop a universal method for large
protein separation online with mass spectrometry. Several problems were observed and solved
such as bubble formation and contamination of the sheath liquid. These problems hampered its
routine use in the laboratory. Improvements in operation and stability that yielded a robust
platform for routine intact protein analysis will be described along with examples of protein
separations using the nanosheath CE/MS. Using this improved interface a divert mechanism is
enabled that allows selective deletion of a peak in the total ion electropherogram and diversion
away from the MS source during capillary flushing. In summary, CE-MS has become a routine
tool in my lab for solving critical problems in protein characterization.
NOTES:
33
Knock! Knock! Who’s there? “Maurice” Who?
SungAe Suhr Park
Amgen Inc., Thousand Oaks, CA USA
Abstract not available at time of print.
NOTES:
34
Now You CE Me, Now You Don’t: The Evolving Role of CE in the Control Strategy
Sarah Kennett
CDER, FDA, Silver Spring, MD USA
The biopharmaceutical world has primarily used a standard control strategy for many years - the
manufacturing process is run under the control of an extensive set of parameters with specific
operating ranges, the product is tested to evaluate numerous quality attributes prior to release of
the product for clinical study use or to the market, and similarly extensive testing is performed to
assess and ensure product stability. Over the last decade, CE methods have become increasingly
popular for use in the background, i.e., for characterization of the product and process, and as
methods that appear on official release and stability specifications for drug substances and drug
products. With the help of increasingly advanced techniques to investigate complex processes
and products and the knowledge derived, industry appears to have reached an inflection point in
setting control strategies, which might result in the disappearance of some CE-based testing from
its current prominent place on specifications. This talk will provide a regulator’s view of the
roles of CE methods in the current and changing/advancing biopharmaceutical space.
NOTES:
35
Application of Microchip Capillary Electrophoresis in the Quality Control Laboratory
Jeff Schneiderheinze
Regeneron Pharmaceuticals, Inc., Tarrytown, NY USA
Capillary electrophoresis is a powerful tool in the Biopharmaceutical industry for the analysis
and characterization of therapeutic protein products. Capillary electrophoresis using sodium
dodecyl sulfate (CE-SDS) has become widely adopted for use in Quality Control (QC) batch
release and stability testing as well as process characterization and validation to gain an in-depth
understanding of product purity and fragmentation. Despite the adaptation of this methodology,
the technology remains somewhat limited in terms of throughput as well as hands on time
required by the analyst. In addition, process sample matrices containing high concentrations of
salt are often not compatible with CE-SDS and therefore require additional sample handling and
manipulation prior to analysis.
In recent years, microchip capillary electrophoresis (MCE) has emerged with the potential of
dramatically reduced sample analysis times and a simplified user interface while maintaining the
robust performance and data reliability required for QC analysis. In this presentation, we will
discuss the development and application of MCE in the QC environment as well as
comparability data between MCE and traditional CE-SDS methodology. In addition, we will
discuss the robustness of MCE as a “platform” technology and compare instrument performance
in the QC laboratory.
NOTES:
36
CE Applications in a QC Environment
Jian Zhang, Kimia Rahimi, David Fischer, Thomas Niedringhaus, David Michels, Sarah Du
Genentech, a Member of the Roche Group, South San Francisco, CA USA
High performance capillary electrophoresis (CE) technologies have been widely and successfully
implemented in the control systems for biotherapeutics such as monoclonal antibodies (mAbs).
At Genentech, with decades of experience applying CE technologies for clinical and commercial
products, platform CE methods were developed, optimized and validated for use in quality
control (QC). These methods are used to monitor certain key product quality attributes of
therapeutic mAbs, such as size variants and charge variants. Platform CE-SDS and iCIEF
methods are routinely used in QC for batch release and stability monitoring. Recent advancement
in instrumentation also enhanced the performance, robustness and user experience in GMP
testing. In this presentation, we will report the experience of CE method validation, transfer, and
trouble shooting in an IMP-QC environment.
NOTES:
37
The NISTmAb: A Comprehensively Characterized Reference Material to Support
Biopharmaceutical Analytical Technology Development
Abigail Turner1, John Schiel2
1Institute for Bioscience and Biotechnology Research (IBBR), Rockville, MD USA, NIST,
Gaithersburg, MD USA
The National Institute of Standards and Technology (NIST) has recently developed the
NISTmAb Reference Material (RM 8671), an IgG1κ class-representative monoclonal antibody.
The NISTmAb is intended to serve as a platform for harmonization and open innovation in the
biopharmaceutical analysis community. It is a widely and longitudinally available test material
that will support novel technology and method development, serve as an external system
suitability control, and provide a medium for open access information sharing. Here, we provide
an overview of the NISTmAb program, with special emphasis on NISTmAb analysis by
capillary electrophoresis methods including CE-SDS, cIEF, CZE, and CE-ESI-MS2. We
demonstrate comprehensive electrophoretic characterization of the NISTmAb reference material
and discuss its utility as a reference standard in CE assay development.
NOTES:
38
CE Analysis in GMP Environment – What is that Noise?
Jiu-Li Song, Lan Li
Amgen Inc., Thousand Oaks, CA USA
Limit of Detection (LOD) and Limit of Quantitation (LOQ) are important characteristics of an
analytical method. Determination of LOD and LOQ directly impacts the reported analytical
results. One way to determine the LOD or LOQ is based on the signal-to-noise ratio, which has
been used for many years as the strategy for Capillary Electrophoresis (CE) or Liquid
Chromatography based assays. Therefore, noise determination plays an important role in result
accuracy and consistency in the GMP environment. We compared different ways to determine
noise within 32 KaratTM software and in other software such as EmpowerTM and ChromeleonTM.
We also evaluated the noise determination by varying data acquisition rate, varying the time
frame and segment for noise calculation, comparing injection to injection and run to run
variation. The LOQ obtained from noise was further verified by analyzing a monoclonal
antibody near the LOQ level with desired precision and accuracy. This study provided insight on
selecting appropriate noise calculation to determine LOD/LOQ and an understanding of the
impact to LOD/LOQ when data are imported to a different chromatography data system for
processing.
NOTES:
39
Development of an icIEF Method for a Poorly Soluble ADC Derived from an IgG2 mAb
Conjugated with a Charged Drug-linker
Adam Fung, Lily Liu
Agensys, Inc., Santa Monica, CA USA
Imaged capillary isoelectric focusing (icIEF) is commonly used to monitor the charge variant
distributions of monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) for release
and stability testing. Method development can be especially challenging for complex molecules
such as ADCs, and particularly those ADCs conjugated with charged drug-linkers. Some
challenges in the development and characterization of an icIEF method used to monitor the
charge variant distribution of an ADC derived from an IgG2 monoclonal antibody conjugated
with a charged drug-linker will be highlighted. We also introduce a unique approach to
improving solubility and stability of a highly sensitive ADC that is prone to aggregation under
common icIEF matrices and conditions.
NOTES:
40
Microfluidic Capillary Electrophoresis-Electrospray Devices for Analysis of
Biopharmaceutical Materials
J. Michael Ramsey
University of North Carolina, Chapel Hill, Chapel Hill, NC USA
We have pioneered the development of a sensitive, stable, and efficient microchip electrospray
interface that enables the integration of MS detection with rapid and highly efficient microfluidic
separation methods. The separative performance of these devices is near the theoretical
diffusional limit for cationic species. This performance is achieved through the use of novel
surface modification strategies that result in highly homogeneous surface characteristics. These
devices yield electrospray ionization (ESI) performance commensurate with commercial
nanoESI emitters without sacrificing the quality of microfluidic separations. Compared to CE-
MS performed using fused silica capillaries, microchip CE-MS can achieve greater separation
efficiency in shorter analysis times as the integrated injection and ESI functional elements
greatly reduce extra-column band broadening. Microchip CE-ESI-MS has been used for
challenging applications such as the characterization of intact biopharmaceuticals and antibody-
drug conjugates, where achieving optimal separation efficiency is crucial for the success of the
analysis. Moreover, peptide mapping can also be performed rapidly with high coverage. Most
of these separations are completed in less than three minutes and most samples require minimal
sample preparation.
NOTES:
41
Cutting-Edge Mass Spectrometry for mAbs & Related Product Structural
Characterization
Elsa Wagner-Rousset
Centre d'Immunologie Pierre Fabre, Saint Julien en Genevois, France
Monoclonal antibodies (mAbs) are highly complex tetrameric glycoproteins that must be
extensively analytically and structurally characterized to become drug candidates. This is also
true for biosimilar and biobetter antibodies and for IgG-related products such as antibody drug
conjugates (ADCs). These immunoconjugates are based on highly cytotoxic Small Molecular
Drugs (SMDs) covalently attached via conditionally stable linkers to mAbs and are among the
most promising next generation empowered biologics for cancer treatment. ADCs are more
complex than naked mAbs, as the heterogeneity of the conjugates adds to the inherent
microvariability of the biomolecules. The development and optimization of ADCs rely on
improving their analytical and bioanalytical characterization by assessing several critical quality
attributes (CQAs), namely the distribution and position of the drug, the amount of naked
antibody, the average drug to antibody ratio (DAR), and the residual drug-linker and related
product proportions (SMDs). Progresses of multi-level (top, middle, bottom, small molecular
drugs) state-of the art mass spectrometry methods (Native MS, Ion Mobility-MS, CESI-MS, 2D-
LC-MS, Extended Bottom Up, Top Down Sequencing) combined with chromatographic and
electrophoretic techniques will be presented. They will be illustrated by extensive structural
characterization of reference FDA and EMA approved therapeutic mAbs and ADCs.
Wagner-Rousset E. et al. Fast workflow to rank critical versus non-critical charge
variants of therapeutic antibodies. Journal of Chromatography A, 2016 in press.
Beck A. et al Cutting-Edge MS methods for multi-level ADC structural characterization.
Expert Review of Proteomics, 2016.
Resenman A, Beck A et al. Full Validation of Therapeutic Antibody Sequences by
Middle-Up Mass Measurements and Middle-Down Protein Sequencing. mAbs, 2016.
Gahoual R, Beck A et al. Independent highly sensitive characterization of asparagine
deamidation and aspartic acid isomerization characterization by sheathless CZE-ESI-
MS/MS. J Mass Spec 2016.
François YN, Beck A et al. Characterization of cetuximab Fc/2 dimers by off-line CZE-
MS. Anal Chem Acta 2016.
Gahoual R. Cutting Edge capillary electrophoresis characterization of monoclonal
antibodies and related products. J. Chrom. B 2016.
NOTES:
42
CE-MS Analysis of A Single Dose Pharmacokinetic Study of Two Fc-Fusion Protein
Constructs
Mei Han
Amgen Inc., South San Francisco, CA USA
Abstract not available at the time of print.
NOTES:
43
NOTES:
44
Workshop I: Troubleshooting
Monday, September 26
17:00-18:00
Crystal Ballroom
Facilitators:
Tim Blanc, Eli Lilly and Company, Branchburg, NJ USA
Cari Sänger-van de Griend, Kantisto BV, Baarn, Netherlands
Scribe:
Tara Enda, Eli Lilly and Company, Branchburg, NJ USA
Henry Luo, Regeneron Pharmaceuticals, Inc., Tarrytown, NY USA
Joel Welch, CDER, FDA, Silver Spring, MD USA
Analytical methods subject matter experts (SMEs) play an important business role by ensuring
the success of technologies in labs supporting characterization and GMP testing. As a
community of Capillary Electrophoresis SMEs, sharing expertise among the industry is one of
the primary objectives of the CE Pharm Meeting and is the focus in our annual troubleshooting
workshop. Each laboratory makes unique distinctions about common problems and devise
unique and cleaver solutions to such problems within their organization. Some have
affectionately coined this acquired information as “Tribal Knowledge.” Internally, it may be
viewed as too trivial to publish, even though it is critical to the performance of important
methods. The goal of this workshop is to share and harness such tribal knowledge across our CE
community.
While lively and informative discussions are the goal, a picture (or Electropherogram) can
provide a much higher level of clarity to the discussion. This year we have invited attendees to
submit electropherograms representative of their troubleshooting issues. The hope is that the
electropherograms will bring a new level of clarity to questions that focus discussion and send
attendees home with solutions. Two years ago we began soliciting examples for this workshop
and the response has been impressive. A report of last year’s workshop can be found at
http://www.casss.org/page/CEPharmTroubleshoot. Also a report of this year’s troubleshooting
session will be published at the CASSS website.
45
NOTES:
46
Round Robin Table Discussions
Tuesday, September 27, 2016
16:50 – 17:50
There are 13 roundtable topics. The plan is for these to be active discussions, not presentations
or lectures. To create useful discussion we are going to try and limit each topic to 10 attendees.
Seating will be on a first come, first serve basis. These discussions will include a facilitator,
whose role is to help assist the discussion and ensure a lively exchange, and a scribe, whose role
is to make general, anonymous notes about the discussion that will be posted on the CE Pharm
2016 website.
Listed below is a quick view of the Roundtable Topics, Facilitators and Scribes:
Table 1 CE-SDS: Briding Between Different Kits. Reduced vs. Non-reduced - When
to Focus on One?
Sarah Kennett, CDER, FDA
David Michels, Genentech, a Member of the Roche Group
Table 2 CE-SDS: Peak Identification
Christian Neusüß, Aalen University
Göran Hübner, Boehringer Ingelheim Pharma GmbH & Co. KG
Table 3 What are Opportunities and Challenges for Further Implementation of CE-
MS in Development of Biologics?
Michael Knierman, Eli Lilly and Company
Nathan Lacher, Pfizer, Inc.
Table 4 Chip Based Separations vs Classical CE Separations
Friederike Winkhaus, Roche Diagnostics GmbH
SungAe Suhr Park, Amgen Inc.
Table 5 Glycan Analysis by HILIC-LC or CE. Are you Doing One or Both?
Sherry Guo, Genentech, a Member of the Roche Group
C. Mark Lies, SCIEX
47
Round Robin Table Discussions
Table 6 CE's Role for Challenging Analytical Problems: New Therapeutic Protein
Formats like Fusion Proteins, ADCs, Subvisible Particles, etc.
Lars Geurink, Janssen Infectious Diseases and Vaccines
Cari Sänger - van de Griend, Kantisto BV
Table 7 With Minimal Manufacturing Experience, What are Some Strategies for
Setting Quantitative Specifications for Charge Variants and Size Variants?
Richard Rustandi, Merck & Co., Inc.
Hermann Wätzig, TU Braunschweig
Table 8 Affinity Capillary Electrophoresis
Kathir Muthusamy, Regeneron Pharmaceuticals, Inc.
Mona Mozafari, TU Braunschweig
Table 9 What are the Common Strategies for Introducing New Instruments (i.e. Lab
Chip, Maurice etc.) into the a GMP Testing Environment?
Xin Jiang, ProteinSimple
Zoran Sosic, Biogen
Table 10 Do people Rely on Release Methods for Late Phase Process Characterization
or Have High Throughput Workflows in Place to Support Such Activities?
How Similar do the Product Quality have to be Between the Release and the
High Throughput Method?
Nomalie Jaya, Seattle Genetics, Inc.
Mei Han, Amgen Inc.
Table 11 QbD and DoE in CE: Nice-to-have or Need-to-have?
Peter Bryngelson, Biogen
Joel Welch, CDER, FDA
Table 12 Adsorption and Capillary Conditioning: Bare fs vs. Capillary Coating(s)
Ewoud van Tricht, Janssen Infectious Diseases and Vaccines
Gordon Freckleton, Eli Lilly and Company
Table 13 Instrument Quality, Reliability and Failure Rate
Jason Matthews, Biogen
Tim Blanc, Eli Lilly and Company
48
Technical Seminars Technical Seminar/Lunch and Learn
Monday, September 26
12:25 – 13:25
Crystal Ballroom
Run Your Assays Faster: Getting the Quantitative Answers You Need For N-Glycan, CE-
SDS, and CZE Applications Without the Wait
Matt Salem
SCIEX, Brea, CA USA
Comprehensive characterization of protein therapeutics through highly quantitative data is
crucial for the biopharmaceutical industry. More importantly, the ability to gather high resolution
data while simultaneously shortening the overall analysis time has proven vital to both
biopharmaceutical companies and CROs. With the introduction of the Fast Glycan Labeling and
Analysis Kit from SCIEX, you can run glycans up to 10x faster than LC! Performing glycan
sample prep, analysis and ID on up to 96 samples can now be done in a single day. CE-SDS
analysis for molecular weight or purity determination has also been revamped to maintain 3 logs
of dynamic range (with UV detection) while cutting sample analysis time in half by utilizing the
High Speed CE-SDS method. Lastly, Capillary Zone Electrophoresis (CZE) can be utilized to
get charge heterogeneity information in just a fraction of the time it takes to run traditional cIEF
assays. These core biologics characterization applications have been optimized so that you can
get to the answers you need, faster.
NOTES:
49
Technical Seminar/Lunch and Learn
Tuesday, September 27
11:45 – 12:45
Crystal Ballroom
Meet Maurice, One-stop cIEF and CE-SDS for your Biologics
Scott Mack, Irina Kazakova, Jiaqi Wu, Xin Jiang
ProteinSimple, San Jose, CA, USA
Analysis of a therapeutic protein by cIEF and CE-SDS is critical in establishing a product’s
identity, purity, and heterogeneity. Performing these separations traditionally requires the
implementation of both imaged and fixed window CE systems. Maurice, a next generation
analytical platform, combines both detection schemes into a single instrument, allowing cIEF or
CE-SDS data to be generated in a snap. New ready-to-go cartridge designs and automated clean
up procedures dramatically reduces system maintenance while eliminating cross-contamination
concerns. Complimenting Maurice’s streamline operation is a novel native fluorescence cIEF
detection mode capable of increasing sensitivity 3 to 5 fold over absorbance.
A comprehensive overview of the Maurice instrument family will be presented. The subjects
that will be covered include an introduction into the system’s features, insights into instrument
development, and the results from internal evaluations. Charge and size data from multiple
compounds will provide for an accurate estimation of system performance and comparability to
currently employed CE instrumentation.
Also join us in a round-table discussion with the early users of the Maurice system, and experts
from ProteinSimple, to hear about the first-hand evaluation of Maurice, and our ongoing KOL
program.
NOTES:
50
Technical Seminar/Lunch and Learn
Wednesday, September 28
12:30 – 13:10
Diamond Room
High Throughput and High Resolution N-glycan Analysis Using Multicapillary CE and
Novel Fluorescent Dyes
Shaheer Khan
Thermo Fisher Scientific, South San Francisco, CA USA
Glycosylation is one of the key critical quality attributes of mAb based biotherapeutics.
Glycosylation changes can impact biological drug’s safety, efficacy, clearance and
immunogenicity, making it necessary to accurately detect changes. Glycan profiling begins at
cell line development and continues through process development. Current glycan analysis
methods involve laborious multistep sample preparation that takes anywhere from a day to
multiple days for 96 samples, followed by single channel LC or CE separation.
Here, we report the development of a high throughput glycan analysis method utilizing a 24
capillary polymer filled array with laser induced fluorescence detection. For glycan labeling
along with conventional APTS dye, two new rapidly reacting fluorescent dyes were developed.
Glycan cleavage, dye labeling and excess dye removal steps were streamlined for automation.
We also eliminated the toxic sodium cyanoborohydride chemistry and vacuum centrifugation
steps. Our new analysis method can analyze glycans from low glycoprotein input (< 10ug) and
provide glycan quantitation data from 96 samples in 7-9hrs with <3 hours of hands-on time.
Some of the IgG glycans that were unresolved when labeled with APTS were fully baseline
resolved when labeled with our proprietary dyes. These novel dyes on a simple streamlined
workflow in combination with a multi capillary CE instrument offer a high resolution high
throughput glycan analysis platform for rapid separation and quantitation of antibody glycans.
Our fully integrated solution of easy sample prep, high throughput instrumentation and software
can address the current unmet need of a high throughput and high resolution glycan analysis
solution.
NOTES:
51
NOTES:
52
Poster Abstracts
Deep Dive into CE (Next 10 years)
P-101
Multiple Modes Application of Capillary Electrophoresis in Aptamers Selection Qu Feng
Beijing Institute of Technology, Beijing, China
P-102
High Resolution CE-MS Analysis of APTS-labeled Monoclonal Antibody N-glycans Andras Guttman1, Marton Szigeti2, Bryan Fonslow3 1University of Debrecen, Debrecen, Hungary, 2University of Pannonia, Veszprem, Hungary, 3TSRI, La Jolla, CA USA
P-103
Simplifying the Process Data Interpretation for Identifying and Assigning Glycan
Structures Andras Guttman1, Gabor Jarvas1, Marton Szigeti2, Jeff Chapman3 1University of Debrecen, Debrecen, Hungary, 2University of Pannonia, Veszprem, Hungary, 3SCIEX, Brea, CA USA
P-104
Potential Interferences of Ampholytes to Imaged Capillary Electrophoresis (iCE) Testing
of mAb and ADC Xiaoping He1, Jianming Jim Mo2 1Pfizer, Inc., St Louis, MO USA, 2Pfizer, Inc., Chesterfield, MO USA
P-105
A Novel Nanospray Liquid Junction Interface for Versatile CE-MS Jana Krenkova1, Rob Haselberg2, Govert W. Somsen2, Frantisek Foret1 1Institute of Analytical Chemistry, v. v. i., Brno, Czech Republic, 2Vrije Universiteit Amsterdam,
Amsterdam, Netherlands
53
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Method Lifecycle Development
P-106
Method Development of an icIEF Method for the Replacement of a Pre-cast IEF Gel
System within an Analytical Method Lifecycle Franziska Hübner, Frederic Zoller
Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany
P-107
Validating a New Quantitative CE method for a Marketed Recombinant Protein Product
to Support Control System Updates Liang Jia
Genentech, a Member of the Roche Group, South San Francisco, CA USA
P-108
Development and Evaluation of Fast Charge Heterogeneity Methods for Monoclonal
Antibodies and New Modalities Maria Eleanor Le, Jeremy Primack, Lan Li, SungAe Suhr Park, Tzu-Chin Chen, Chao-Hsiang
Wu
Amgen Inc., Thousand Oaks, CA USA
P-109
Implementation of Capillary Electrophoresis for Viral Vaccine Development Support in a
GMP Environment Lars Geurink1, Ewoud van Tricht1, Cari Sänger – van de Griend2 1Janssen Infectious Disease and Vaccines, Leiden, Netherlands, 2Kantisto BV, Baarn,
Netherlands
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56
Novel Technologies (Next 5 years)
P-110
Flow Induced Dispersion Analysis (FIDA) as a Novel Technology for Protein
Quantification in Plasma and for Probing Immune Responses in Patients Nicklas N. Poulsen1, Morten E. Pedersen1, Jesper Østergaard1,2, Nickolaj J. Petersen1, Niels H.
H. Heegaard3, Henrik Jensen1,2 1University of Copenhagen, Copenhagen, Denmark, 2FIDA-Tech Aps, Roskilde, Denmark, 3Statens Serum Institut, Copenhagen, Denmark
P-111
Flow Induced Dispersion Analysis for Probing non-Covalent Interactions and Quantifying
Large Biomolecules: A New Approach to Ligand Binding Assays Nicklas N. Poulsen1, Jesper Østergaard1,2, Nickolaj J. Petersen1, Henrik Jensen1,2 1University of Copenhagen, Copenhagen, Denmark, 2FIDA-Tech Aps, Roskilde, Denmark
P-112
Automated cIEF Sample Preparation by Maurice On-Board Mixing Jessica Dermody, Lekha Priya, Annegret Boge, Scott Mack
ProteinSimple, San Jose, CA USA
P-113
Application of Optimized Microchip-Based Electrophoresis for Monoclonal Antibody
Product Quality Analysis Ya Fu, Jun Xu, Ming Zeng, Tapan Das
Bristol-Myers Squibb Company, New Brunswick, NJ USA
P-115
Evaluation of a Novel Instrumentation for Capillary Electrophoresis-Sodium Dodecyl
Sulfate (CE-SDS) Analysis for Antibody-Drug Conjugates Natalie Jones, Nomalie Jaya, Oscar Salas-Solano
Seattle Genetics, Inc., Bothell, WA USA
P-116
An Integrated System for High-throughput, User-friendly N-Glycan Analysis Using Rapid
Separation by Capillary Electrophoresis Michael Kimzey, Andres Guerrero, Aled Jones, John Yan, Zoltan Szabo, Shirley Ng, Alexander
Gyenes, Justin Hyche, Emily Dale, Ted Haxo, Sergey Vlasenko
ProZyme, Hayward, CA USA
57
P-117
Optimization of Maurice CE-SDS Performance: Effect of Sample Composition and
Preparation Irina Kazakova, Annegret Boge, Jessica Dermody
ProteinSimple, San Jose, CA USA
P-118
NGS Library Validation by an Automated Capillary Gel Electrophoresis Alice Lin, Jemmie Chang, Eric Tsai, Varouj Amirkhanian
BiOptic Inc., Taipei, Taiwan
P-119
Detection of Microsatellite Instability in Colorectal Cancer by an Automated Capillary Gel
Electrophoresis Alice Lin, Varouj Amirkhanian, Eric Tsai
BiOptic Inc., Taipei, Taiwan
P-120
Rapid and Simple Sample Preparation for High Throughput and High Resolution Glycan
Analysis by Capillary Electrophoresis Jenkuei Liu, Shaheer Khan, Bharti Solanki
Thermo Fisher Scientific, South San Francisco, CA USA
P-121
cIEF and CE-SDS Characterization of NISTmAb by Maurice Scott Mack, Irina Kazakova, Annegret Boge, Jessica Dermody
ProteinSimple, San Jose, CA USA
P-122
Real-time Assay Optimization on a Microfluidic Platform for Pharmaceutical Analysis Anubhav Tripathi1, David Weinberger1, Jingjing Wong2, Derek Troiano2, I-Jane Chen2, Wael
Yared2, Jim Hudson2, Laurel Provencher2 1Brown University, Providence, RI USA, 2PerkinElmer, Hopkinton, MA USA
P-123
Multilevel Characterization of Monoclonal Antibodies using Microfluidic CE-MS
Technology Erin Redman, Joshua Guerrette, Scott Mellors
908 Devices, Inc., Carrboro, NC USA
58
P-124
Evaluating the Imaged Capillary Isoelectric Focusing (icIEF) Capabilities of a Novel
Instrumentation for Antibody-Drug Conjugates Kevin Strozyk, Nomalie Jaya, Oscar Salas-Solano
Seattle Genetics, Inc., Bothell, WA USA
P-125
Evaluation of Microchip HT Capillary Electrophorsis Assays for QC testing - a Multi-site
Ring Trial Friederike Winkhaus
Roche Diagnostics GmbH, Penzberg, Germany
P-126
Capillary Isoelectric Focusing with Intrinsic Fluorescence Whole-Column Detection Jiaqi Wu
ProteinSimple, Toronto, ON Canada
59
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Protein Analysis and Emerging Therapeutics
P-127
Optimization of Denaturation Conditions in a Reducing Microfluidic Capillary Gel
Electrophoresis Purity Assay for an IgG1 Monoclonal Antibody Troy Adams
GlaxoSmithKline, King of Prussia, PA USA
P-128
High Sensitivity CZE-ESI-MS Investigations and Applications Emily Amenson1, Liangliang Sun2, Norman Dovichi1 1University of Notre Dame, Notre Dame, IN USA, 2Michigan State University, East Lansing, MI
USA
P-129
Comparative mAb Charge Heterogeneity Assays by CESI-MS and CZE/cIEF-UV Analyses Steve Lock1, Edna Betgovargez2, Bryan Fonslow3, Ying Zhang4, Olga Friese4, K. Steven Cook4,
John Yates III5, Jason Rouse6 1SCIEX, Warrington, UK, 2SCIEX, Brea, CA USA, 3SCIEX Separations, San Diego, CA USA, 4Pfizer, Inc., Chesterfield, MO USA, 5The Scripps Research Institute, La Jolla, CA USA, 6Pfizer,
Inc., Andover, MA USA
P-130
A Pretreatment Approach to Characterize Complex Fc-based Fusion Proteins through CE-
SDS Paras Bhatia
Takeda Pharmaceuticals, Cambridge, MA USA
P-131
Method Development and Characterization of Fusion Protein using Imaged Capillary
Isoelectric focusing (iCIEF) Ying-Chen Chen, Anulfo Valdez, Girija Krishnamurthy
Bristol-Myers Squibb Company, Bloomsbury, NJ USA
P-132
Targeting Desired N-Glycosylation Profiles Through The Use Of Glycosylation Enhancing
Feeds and High Throughput and High Resolution N-Glycan Analysis by Multi Capillary
Electrophoresis Shaheer Khan1, Jaime Goldfuss2, Shawn Barrett2, Mark Stramaglia2, Bharti Solanki-Nand1,
Johnie Young1, Jenkuei Liu1, Baburaj Kunnummal1, Steve Gorfien2 1Thermo Fisher Scientific, South San Francisco, CA USA, 2Thermo Fisher Scientific, Grand
Island, NE USA
61
P-133
Evaluation of the ProteinSimple Maurice System for Reduced and Non-reduced CE-SDS
Applications Mee Ko, SungAe Park, Lan Li, Jeremy Primack, Tzu-Chin Chen, Chao-Hsiang Wu
Amgen Inc., Thousand Oaks, CA USA
P-134
Effects of Denaturants and Stabilizers in the Analysis of Charge Variants of Monoclonal
Antibodies using Imaged Capillary Isoelectric Focusing Wenkui Lan, Mary Krause, Ronak Shah, Kathleen Kelly, Mark Bolgar, Rajesh Gandhi
Bristol-Myers Squibb Company, New Brunswick, NJ USA
P-135
Why Light and Peroxide Stress Cause Increase in Acidic Species and Decrease in Protein
Unfolding Temperature Ning Li, Dipali Patel, Christopher Shaw, Anil Kumar Meda Kavadi, June Kim, Luke Bergerud,
Derek Schildt
AstraZeneca, Frederick, MD USA
P-136
Capillary Gel Electrophoresis as a Tool for Purity Profiling of Bispecific Monoclonal
Antibodies Jasna Maksimoska
Janssen Pharmaceutical R&D, LLC, Malvern, PA USA
P-137
Development of an Imaged Capillary Isoelectric Focusing (icIEF) Denatured and Reduced
Method to Analyze Charge Heterogeneity of a Multi-domain Complex Glycoprotein Janette Mathis, Byron Kneller, Michelle A. Emrick
CMC Biologics, Bothell, WA USA
P-138
Innovator and Biosimilar Monoclonal Antibody-Peptide Map Analysis Using Capillary
Electrophoresis and Comparison Using Chromatograph Comparison Software Arunkumar Padmanaban1, Michael Yap2 1Agilent Technologies India Pvt Ltd, Bangalore, India, 2Agilent Technologies, Santa Clara, CA
USA
P-139
A Platform DoE Approach for Non-Reducing CE-SDS Method Development for mAbs Irina Perdivara, Clara Smith, Greg Adams
FUJIFILM Diosynth Biotechnologies, Morrisville, NC USA
62
P-140
Complementarity Study of the N-linked Glycosylation of a Development-Phase
Therapeutic Glycoprotein Laura Salmeron1, Andras Guttman2, Anghel Jimenez1, Shiping Fang1 1Halozyme Therapeutics, San Diego, CA USA, 2University of Debrecen, Debrecen, Hungary
P-141
Analytical Characterization of Protein-Polymer Conjugates Used for Long Acting Delivery
Ocular Therapeutics Cinzia Stella
Genentech, a Member of the Roche Group, South San Francisco, CA USA
P-142
Novel CZE Method for Quantification of Intact Virus Particles in Complex Matrices –
Quality by Design method development Ewoud van Tricht1, Lars Geurink1, Cari Sänger – van de Griend2 1Janssen Infectious Diseases and Vaccines, Leiden, Netherlands, 2Kantisto BV, Baarn,
Netherlands
P-143
Quantitative CE-MS Glycomics Using Twoplex Stable Isotope Labelling Csaba Varadi, Stefan Mittermayr, Silvia Millán-Martín, Jonathan Bones
NIBRT, Dublin, Ireland
P-144
The Effect of Histidine Formulation Buffer on the Profile and Apparent Isoelectric Point of
Therapeutic Proteins Analyzed by Imaged Capillary Isoelectric Focusing Chao-Yu Chen, Edward Wang, Brad Hayes
OncoMed Pharmaceuticals, Redwood City, CA USA
P-145
A Theoretical Model for Correlation of Non-Glycosylated Heavy Chain Detected by
Conventional CESDS and Micro-Chip Based Electrophoresis Jun Xu, Ya Fu, Ming Zeng, Tapan Das
Bristol-Myers Squibb Company, New Brunswick, NJ USA
P-146
Middle-down Proteolysis with Imaged Capillary Isoelectric Focusing and Capillary Gel
Electrophoresis for Domain-specific Characterization of Innovator and Generic Rituximab Zichuan Zhang, Julia Ding, Ronel Perrault, Yun Zhao
PPD, Middleton, WI USA
63
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64
Regulatory
P-147
Examination of Interferon Alpha-2 Charged Variants and Assessment of the Active
Ingredients Integrity in Finished Products using Capillary Electrophoresis and High
Performance Liquid Chromatography Simon Sauve
Health Canada, Ottawa, ON Canada
P-148
Standard Test Methods Based on Capillary Electrophoresis Xinying Zhao
Beijing Centre for Physical & Chemical Analysis, Beijing, China
65
Small Molecule
P-149
Exploring the Capabilities of Capillary Electrophoresis in Solving Difficult Separations for
Small and Large Molecules Van Truong
Merck & Co., Inc., Rahway, NJ USA
P-150
Ultrasensitive Determination of 5-methylcytosine and 5-hydroxymethylcytosine in Genomic
DNA by Sheathless Interfaced CE-MS Fang Yuan1, Xiao-Hui Zhang1, Ji Nie1, Hong-Xu Chen2, Ying-Lin Zhou1, Xin-Xiang Zhang1 1Peking University, Beijing, China, 2Sciex Co (Shanghai), Beijing, China
66
Young Scientist
P-151
Simultaneous Determination of Protein Affinity and Heterogeneity by Capillary
Electrophoresis–mass spectrometry
Rob Haselberg1, Elena Domínguez Vega1, Gerhardus J. de Jong2, Govert W. Somsen1 1Vrije Universiteit Amsterdam, Amsterdam, Netherlands, 2Utrecht University, Utrecht,
Netherlands
P-152
Investigating the Concentration-dependant Binding of Heparinoids to Albumins using
Affinity Capillary Electrophoresis Mona Mozafari
TU Braunschweig, Braunschweig, Germany
P-153
Affinity Capillary Electrophoretic and Computational Methods for Binding Studies of P-
selectin with Heparinoids Mona Mozafari
TU Braunschweig, Braunschweig, Germany
P-154
Capillary Zone Electrophoresis-electrospray Ionization-Mass Spectrometry for Xenopus laevis
Metabolomic Analysis
Nicole M. Schiavone, Jennifer Arceo, Danielle A. Boley, Elizabeth Peuchen, Norman J. Dovichi
University of Notre Dame, Notre Dame, IN USA
P-155
Coupling Capillary Zone Electrophoresis to a Q Exactive HF Mass Spectrometer for Top-
down Proteomics: 580 Proteoform Identifications from Yeast Yimeng Zhao, Liangliang Sun, Guijie Zhu, Norman Dovichi
University of Notre Dame, Notre Dame, IN USA
67
Late Breaking
LB-01
CE-SDS Validation Challenges in Quality Control
Cheryl Lovato, Koman Joe, Jennifer Moore, Lisa Stefanich
Genentech, a Member of the Roche Group, South San Francisco, CA, USA
LB-02
Capillary Electrophoresis – Mass Spectrometry for Top Down Proteomics in Food And
Clinical Studies
Andreas Krupke1, Chien-Hsun Chen2, Shiaw-Min Chen1, Achim Karger1, Steve Williams1,
Michael Wenz1, Daniel Lopez-Ferrer2 and Aran Paulus2 1Thermo Fisher Scientific, South San Francisco, CA USA, 2Thermo Fisher Scientific, San Jose,
CA USA
68
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