CE in the Biotechnology & Pharmaceutical Industries · knowledgeable speakers and workshop leaders, but also on the interactions and open discussions that take place among the attendees.

1

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

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



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


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



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



to Crystal Ballroom for the “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


14:55 – 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


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


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



to Diamond Room for the “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


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


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.

http://www.casss.org/page/CEPharmTroubleshoot

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

NOTES:

54

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


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


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

55

NOTES:

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


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


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


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


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

NOTES:

60

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


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


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


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


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

NOTES:

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

NOTES:

CE in the Biotechnology & Pharmaceutical Industries · knowledgeable speakers and workshop leaders, but also on the interactions and open discussions that take place among the attendees.

Documents