Expression of heterologous proteins presents many challenges and understanding expression systems is key. The 9th annual Optimizing Protein Expression conference delves into protein expression by examining and enhancing expression
systems, including CHO and other mammalian systems, E. coli, yeast and baculovirus. What is the best expression system for expressing your protein of choice? Ease and cost of scale-up must be considered to ensure successful
bottom-line results. Experts will share case studies and disclose data while divulging details of expression systems’ underlying mechanisms. Comparing and contrasting systems will also be featured to increase understanding in the quest for greater
productivity.
Final Agenda
WEDNESDAY, APRIL 10
7:15 am Registration (Commonwealth Hall) and Morning Coffee (Harbor Level)
7:25 - 8:25 PANEL DISCUSSION: Women in Science – Inspired Professional and Personal Stories (Continental breakfast provided) (Waterfront 1&2)
Moderator:
Jennifer S. Chadwick, PhD, Director of Biologic Development, BioAnalytix, Inc.; Co-Chair, Mentors Advisors and Peers Program, Women In Bio, Boston Chapter
Panelists:
Joanna Brewer, PhD, Vice President, Platform Technologies, AdaptImmune
Charlotte A. Russell, MD, DMSc, CMO, Alligator Bioscience
Susan Richards, PhD, Presidential Scientific Fellow, Translational Medicine Early Development, Sanofi R&D
Kristi Sarno, Senior Director, Business Development, Pfenex
8:30 Chairperson’s Opening Remarks
Philippe Billiald, PharmD, PhD, Professor, Biochemistry, University Paris-Sud & Acticor Biotech
8:40 KEYNOTE PRESENTATION: Early Process Development: Challenges and Opportunities for Emerging Therapies
Nicola Beaucamp, PhD, Head, Process Research, Roche Innovation Center Munich, Pharma Research and Early Development, Roche Diagnostics GmbH
A number of new molecules, different from standard antibody structures, have been advanced into clinics by Roche pRED. In order to discover and develop differentiated biologics, Roche’s strategy is based on engineering technologies leading to new
formats and processes which bear several challenges and many opportunities for technical development towards the clinic and beyond.
9:10 Site-Specific Biotinylation of Secreted Proteins in Mammalian Cells in vivo – Made Viable!
Mark Trautwein, PhD, Senior Scientist, Pharmaceuticals R&D, Preclinical Research, Expression Technologies, Bayer AG
Site-specific biotinylation of secretory target proteins is desirable to achieve in vivo during protein expression in mammalian cells. However, conventional techniques like the AviTag suffer from serious losses in expression
yields, preventing their applicability as a generic approach. In this presentation, I will describe how we have optimized a protein tag to solve the problem and achieve this goal.
9:40 SoluPro™, A Groundbreaking E. coli Platform for Next-Generation Antibody
Scaffolds and Protein Therapeutics
Johan Kers PhD, Vice President,
Research, AbSci
SoluPro™, a game-changing E. coli expression platform that eliminates the formation of inclusion bodies, increases plant efficiencies, and drastically reduces COGs and CapEx outlay. Employing proprietary assays to optimize for product titer, function,
and quality, SoluPro™ produces correctly folded, active protein, at groundbreaking titers in less than 3 months. The platform can produce a wide range of complex proteins including full-length antibodies (4 g/L), Fabs (4.4g/L), and insulin (>20g/L)
in 48 hours or less.
10:10 Coffee Break in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
10:15 Women in Science Speed Networking in the Exhibit Hall (Commonwealth Hall)
10:55 Combining Biophysical Analytics with Next-Generation Sequencing for Deep Characterization of mAb Producing CHO Cell Lines
Holger Thie, PhD, Senior
Manager, Technology and Innovation, Biologics Development, Boehringer Ingelheim Pharma GmbH & Co. KG
Differences in gene expression can lead to unpredicted behavior of mAb producing CHO cell lines during bioprocess development. We observed remarkable differences in N-linked glycosylation between two production clones derived from the same CHO cell line.
NGS-based genetic characterization allowed unraveling the genetic regulation behind the differences in N-linked glycosylation. This study shows how NGS helps to gain a deeper understanding of CHO production cell lines.
11:25 Engineering CHO Cell Lines for the Production of Hard-to-Produce Proteins
Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
Using our high-throughput cell line engineering platform, we have engineered CHO cells able to produce therapeutic proteins that have previously not been possible to produce in CHO cells. This approach may result in improved therapeutic proteins with
better biological properties, such as increased half-life, improved activity, etc.
11:55 Importance of Appropriately Glycosylated Species, and Modulation Efforts, to Achieve Desired Post-Translational Modifications in CHO-Derived Monoclonal Antibodies
Gaurav Chauhan, MS, Associate Principal Scientist, BioProcess Development, Merck & Co., Inc.
The glycosylation profile of the monoclonal antibodies has major impact on the efficacy and safety of the drug and is therefore an important parameter to control during protein production. Since glycosylation is an important parameter for IgG function,
several strategies to modify glycosylation profiles of human IgG or to select the most efficient glycoforms have been explored. Further, considerable efforts have been made to understand how these glycoforms can be modulated as desired. I would like
to present on current understandings of such efforts.
12:25 pm New Tools for Screening & Harvesting Solutions for CHO & HEK293 Cells, for both Transient and Stable Cells
Samuel Ellis, Vice President,
Thomson Instrument Company
Evaluation of different transfection tools, product quality, and titer for both CHO and HEK293 cell lines. Data will be presented on techniques and technology that mimic large-scale bioreactors in non-controlled devices from 1mL-3L. Technologies presented
include well plates and culture tube systems with incorporated filtration methodology. A new direct harvesting technique will also be introduced that eliminates centrifugation while maintaining 0.2um sterile filtration. All of these tools will be
presented with case studies from scientists.
12:55 Luncheon Presentation I: Scaling Flexibility with Fed-Batch Expression in 96-Well Plates and Shaken Single-Use Bioreactors
Annie Ngo, Technical Scientist, Kuhner Shaker Inc
Simple and unique, Kuhner Shaken Bioreactors (SB) mimic shake flask conditions for robust and scalable process development. From advanced 96-well fed-batch screening plates to single-use disposables of 2500L w/v, orbital shaking scales easily based on
kLa and mixing time. Shaken processes offer further advantages of low-shear stress and less foaming – especially beneficial for sensitive cell types. Here we present data for small scale controlled-release fed-batch and data from users
of the 3L-12L SB10 bioreactor.
1:25 Luncheon Presentation II: Generation of High-Yielding Stable Pools Expressing Difficult Target Proteins Through Maxcyte’s Transfection System®
Kevin Guay, Associate Scientist, Jounce Therapeutics
The transient transfection of Human Embryonic Kidney (HEK) cells has long been a workhorse for protein production. Often, however, these reagents suffer from poor yields and high amounts of aggregate formation. Furthermore, multiple production runs can
introduce different post-translation modification patters, which could potentially lead to discrepancies in protein activity. We have employed the route of stable-pool generation to mitigate these issues, providing a great tool for programs in all
stages of drug development.
1:55 Session Break
2:10 Chairperson’s Remarks
Nicola Beaucamp, PhD, Head, Process Research, Roche Innovation Center Munich, Pharma Research and Early Development, Roche Diagnostics GmbH
2:15 Generation of Superior Host Cell Lines for Biomanufacturing
Margaret Lai, MS, Investigator II, Novartis Institutes for BioMedical Research
CHO cells are the most widely used host for large-scale production of recombinant therapeutic proteins. Using transcriptomic approaches, we have identified target genes involved in productivity and product quality. Subsequently, a variety of novel parental
CHO cell lines were generated applying cell line engineering techniques as ZFN or TALEN. These novel knockout CHO cell lines are superior in respect to productivity and/or product quality.
2:45 Achieving Predictable Recombinant Gene Expression in CHO Cells Using CRISPR-Mediated Genome Engineering
Nuša Pristovšek, PhD, Postdoctoral Researcher, The Novo Nordisk Foundation Center for Biosustainability (CFB), Technical University of Denmark
Chinese hamster ovary (CHO) cells are one of the major hosts for production of complex therapeutic proteins. Efficient synthetic biology tools are of great interest to improve production in CHO cell factories. Here, our latest development of these tools
will be demonstrated. Together with high-throughput technologies and systems biology approaches, synthetic biology can pave the way toward accelerated generation of desirable CHO cell factories with predicted culture performance.
3:15 Scaling Up and Scaling Out: Pushing the Boundaries of Transient Protein Production
Ian Wilkinson, PhD, CSO, Absolute Antibody, Ltd.
Whilst transient yields have improved drastically in the last decade, scalable systems are time-consuming and costly to implement. Absolute Antibody has developed systems which scale up and scale out protein expression and purification, enabling the rapid
and cost-effective production of milligram to gram quantities of large panels of proteins.
3:45 Refreshment Break in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
4:45 Problem-Solving Breakout Discussions - Click here for details (Commonwealth Hall)
5:45 Networking Reception in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
7:00 End of Day
THURSDAY, APRIL 11
8:00 am Registration (Commonwealth Hall) and Morning Coffee (Harbor Level)
8:30 Chairperson’s Remarks
Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
8:35 Microbial or Mammalian Cells for Expression of Therapeutic Antibody Fragments: Making the Choice with the End in Mind
Philippe
Billiald, PharmD, PhD, Professor, Biochemistry, University Paris-Sud & Acticor Biotech
Because antibody fragments are usually aglycosylated proteins, many host cell expression platforms can be used for production, including bacteria, yeast and mammalian cells. However, all these expression systems are not equivalent in terms of cell line
development, culture time, product quality and cost of production. We will report differences that may have to be considered before pharmaceutical development and moving forward to the clinic.
9:05 Using Non-Coding RNA for Improving Recombinant Protein Expression and Growth from Mammalian Cells and Microorganisms
Joseph Shiloach,
PhD, Director, Biotechnology Core Lab, NIDDK, NIH
Non-coding RNAs, including microRNA and siRNA in eukaryotes, and small RNA in prokaryotes, are regulatory molecules that can affect protein expression through interactions with specific sections of the host mRNA in mammalian cells, and mRNA and proteins
in bacteria. The utilization of mammalian microRNA and siRNA to enhance growth and protein expression from HEK and CHO cells and affecting E. coli metabolism using bacterial small RNA will be presented.
9:35 Sponsored Presentation (Opportunity Available)
9:50 Overcoming Limitations of Conventional Tag Systems – Strep-Tactin®XT Applications
Dennis Karthaus, MSc, IBA Lifesciences
The Strep-Tactin®XT:Twin-Strep-tag®-purification system enables protein purification at high yields and purity under physiological conditions. Providing the highest binding affinity among all affinity tag systems, the technology fulfills the demands
of mammalian expression systems (e.g. Expi) and is well suited for downstream applications like SPR.
10:05 Coffee Break in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
11:05 Rational Engineering of an Improved Secretion Signal for Pichia pastoris
Benjamin Glick, PhD, Professor, Molecular Genetics and Cell Biology, University of Chicago
The yeast Pichia pastoris is widely employed to secrete heterologous proteins. Typically, the N-terminal portion of pre-pro-alpha-factor is used as a secretion signal. This secretion signal promotes post-translational
translocation into the ER, so proteins that fold in the cytosol are poorly secreted. The alpha-factor pro region can also promote aggregation in the ER. We applied cell biological principles to address both issues. The resulting improved secretion
signal confers dramatic benefits.
11:35 Titer Estimation for Quality Control (TEQC) Method: A Practical Approach for Optimal Production of Protein Complexes Using the Baculovirus Expression Vector System
Yuichiro Takagi, PhD, Associate Professor, Department of Biochemistry and Molecular Biology, Indiana University School of Medicine
The baculovirus expression vector system (BEVS) is becoming the method of choice for expression of many eukaryotic proteins and protein complexes. However, what influences the overall production of proteins or protein complexes remains largely unclear.
We developed the Titer Estimation for Quality Control (TEQC) method, which enables researchers to quantitatively optimize protein expressions utilizing BEVS in a highly reproducible fashion.
12:05 pm BryoTechnology: Large-Scale GMP-Manufacturing of Glyco-Designed Proteins with Moss
Andreas Schaaf,
PhD, CSO, Greenovation Biotech GmbH
BryoTechnology, i.e., moss-based production of biopharmaceuticals, has evolved into a GMP manufacturing technology with products already in clinical development. While leveraging the mosses advantages, comparability to mammalian cell-based technologies
was a priority in process development. Today’s moss process relies on the latest single-use technologies and follows the established routines of mammalian cell-based production. Thus, moss-based production fits easily into existing cleanroom
environments and offers rapid changeover and flexible configuration.
12:35 End of Optimizing Protein Expression