Cambridge Healthtech Institute’s 22nd Annual
Recombinant Protein Expression and Production
Enhancing Expression, Performance, and Process
January 21-22, 2020
Part of the Biotherapeutic Expression & Production pipeline
Great strides have been made in the expression, production, and purification of biotherapeutics. However, hurdles remain to improving the quantity and quality of the efficient expression and production of these valuable biomolecules while minimizing time
and cost. There is a greater demand for higher-throughput expression and purification, as well as more flexible expression platforms. Unfortunately, there is no “universal” production system which can guarantee high yields of recombinant
protein, particularly as every biomolecule causes issues in terms of its own expression. Cambridge Healthtech Institute’s 22nd Annual Recombinant Protein Expression and Production conference explores the newest data and
innovations relating to the best choices in hosts/systems, as well as ways to “rescue” existing systems and make them work more effectively to produce the quality and quantity of the desired biotherapeutic.
Final Agenda
TUESDAY, JANUARY 21
1:00 pm Registration (Sapphire West Foyer)
1:30 Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
2:00 Chairperson’s Opening Remarks
Henry Chiou, PhD, Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific
KEYNOTE PRESENTATION
2:05 Gene Editing and Robotics Solve Upstream and Downstream Problems in HIV Vaccine Manufacturing in CHO Cells
Phillip Berman, PhD, Distinguished Research Professor, Biomolecular Engineering, University of California, Santa Cruz
The envelope protein, gp120, is a major component of vaccines designed to prevent HIV infection. However, production of gp120 in CHO has been challenging due to low expression, a complex glycosylation structure, and sensitivity to proteolysis. We have
used gene editing of the MGAT1 and C1s genes and robotic selection to solve all three problems while at the same time improving the structure of glycan dependent epitopes recognized by monoclonal antibodies.
2:45 A Rapid and High-Yielding Plant-Based Production Platform for Biologics
NEW: Albor Dobon-Alonso, PhD, Senior Microbiologist, Leaf Expression Systems Ltd.
Leaf Expression Systems is a CDMO specializing in plant-based production of recombinant proteins for research and biomedical applications using a proprietary, transient expression technology, Hypertrans®. Here we present our eukaryotic
expression platform that offers rapid, scalable production of high-quality biologics at lower development costs. We will highlight activity data of biosimilar (bevacizumab) and diagnostic MAbs that we have produced, together with examples from our
internal VLP, enzyme and cytokine pipeline.
3:15 The Optimization of Recombinant Protein Expression
Rob Burgess, Chief Business Officer, Sino Biological Inc
Sino Biological boasts the world’s largest selection of recombinant proteins and is a leading provider of CRO services including recombinant protein production. An overview of the company’s state-of-the-art technologies for optimizing
protein expression will be given addressing production challenges and key influencing factors and will include several example case studies.
3:30 Microbial Secretion of Complex Biotherapeutic Proteins by ESETEC® - Case Study with Akari Therapeutics
Arndt Dietrich, PhD, Senior Expert DSP Development, BioProcess Development, Wacker Biotech GmbH
The innovative and highly efficient E. coli expression system ESETEC® enables the secretion of native recombinant proteins into the fermentation broth at high titer, thereby greatly simplifying the purification processes. This has recently been demonstrated
in collaboration with Akari Therapeutics, a Biotech company focused on development anti-inflammatory Biopharmaceuticals.
3:45 Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
4:30 Engineering Cells for Heparin and Heparan Sulfate Production
Charles Glass, PhD, President and CSO, TEGA Therapeutics, Inc.
Heparin and heparan sulfate (HS) are polysaccharide chains that serve as cofactors for specific protein binding and therefore have potential therapeutic qualities as drugs or drug targets. Protein binding specificity is determined by sulfate modifications
along the polysaccharide backbone, which are cell type specific and which can be engineered through genes in the heparin/HS biosynthetic pathway. Through cell engineering, TEGA is testing a library of heparin/HS compositions for therapeutic applications.
5:00 Scalable Production and Purification of Single-Stranded DNA for Therapeutic Nucleic Acid Delivery
Mark Bathe, PhD, Associate Professor, Biological Engineering, Massachusetts Institute of Technology
Viral-like structured DNA and RNA assemblies, also known as DNA or RNA origami, offer the ability to co-formulate gene-length single-stranded DNA or RNA with CRISPR-RNPs, siRNAs, or ASOs, with integration of active cellular targeting and uptake moieties
including sugars and peptides. Scalable bacterial production of custom length and sequence single-stranded DNA will be presented together with co-formulation of siRNA and CRISPR therapeutics with pre-clinical in vivo biodistribution and
toxicity studies in mouse that offer a platform for next-generation nucleic acid therapeutic delivery.
5:30 Close of Day
5:30 - 5:45 Short Course Registration
(Sapphire West Foyer)
5:45 - 8:45 Recommended Dinner Short Course*
SC6: Assembling an Effective Toolbox of Expression Systems to Support Your Drug Discovery Efforts - Detailed Agenda
Instructors:
Richard Altman, Field Application Scientist, Protein Expression, Biosciences Division, Life Sciences Solutions Group, Thermo Fisher Scientific
Henry Chiou, PhD, Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific
Dominic Esposito, PhD, Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research
*Separate registration required
WEDNESDAY, JANUARY 22
7:45 am Registration (Sapphire West Foyer) and Morning Coffee (Sapphire West & Aqua West Foyer)
8:15 Chairperson’s Remarks
Phillip Berman, PhD, Distinguished Research Professor, Biomolecular Engineering, University of California, Santa Cruz
FEATURED PRESENTATION
8:20 Analysis of the Immunoglobulin G (IgG) Secretion Efficiency in Recombinant Chinese Hamster Ovary (CHO) Cells
Takeshi Omasa, PhD, Professor, Department of Material and Life Science, Graduate School of Engineering, Osaka University
In this study, a CHO cell line producing IgG1, which was genetically fused with fluorescent proteins, was established to directly analyze intracellular secretion. The relationship between the amount of intracellular and secreted IgG was analyzed.
An immunofluorescent microscopy study showed that the established clones could be used to analyze the intracellular secretion bottleneck. Also, to improve the production of various therapeutic antibodies, we analyzed the protein secretion
process in CHO cells producing the DTE IgG infliximab.
8:50 Novel Prediction Tool to Evaluate Synonymous Changes in Recombinant Therapeutic Proteins
Nobuko Katagiri, PhD, Research Biologist, Division of Plasma Protein Therapeutics, Office of Tissues and Advanced Therapies, CBER, FDA
Accurate prediction of phenotype based on the genotype is a fundamental goal in the medical genetics field. In silico tools to predict the effects of single missense variations are available from various sources. On the contrary, prediction
of synonymous or multiple mutations remains highly challenging and yet valuable because these sequence changes are often seen in recombinant therapeutics. Our recent efforts towards improvement of such tools will be discussed.
9:20 Boosting VHH Expression Using UNLOCK PICHIA
Iskandar Dib, Principal Scientist Process Development & Analytics, VALIDOGEN GmbH
VHHs, single-domain antibodies derived from camelid species represent the third generation of antibodies and are currently developed for an increasing number of different applications.
Efficient production of VHHs is enabled by VALIDOGEN’s yield-enhancing protein production toolbox known as UNLOCK PICHIA comprising a broad diversity of molecular tools and expression strategies for Pichia. We demonstrate significant
enhancement of VHH expression by a set of helper factors acting along the way from transcription to secretion.
9:50 Coffee Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
10:35 Recombinant Protein Production in Research at Novo Nordisk
Brian Vandahl, PhD, Corporate Vice President, Recombinant Technologies, Global Research Technologies, Novo Nordisk A/S
A discussion of challenges and opportunities in recombinant protein production seen from a Novo Nordisk Research perspective.
11:05 Recent Advances in Screening and Production of Recombinant Proteins in Plants and Plant Cells
Johannes Buyel, Dr rer nat, Dr-Ing, Head, Bioprocess Engineering, Fraunhofer Institute for Molecular Biology and Applied Ecology IME
High-throughput screening systems in 96-well plate format have been developed for plant-based expression. This can facilitate the production of novel and innovative protein biopharmaceuticals that can otherwise be difficult to express in mammalian
or microbial hosts due to toxicity or complexity respectively. We also discuss the translation from screening to production for plant-based systems.
11:35 Antibody Production Using an Inducible CHO Cell Line: Process Development and Intensification
Olivier Henry, PhD, Associate Professor, Chemical Engineering, Polytechnique Montréal
Mammalian-inducible expression systems are increasingly available and offer an attractive platform to produce recombinant proteins. Inducible systems allow to uncouple the growth and the production phases. We have conducted process development
for a cumate-inducible GS-CHO cell line expressing rituximab. We have explored the use of fed-batch and continuous perfusion strategies applied during the pre-induction phase to enhance process performance in terms of product yield and
quality.
12:05 pm Session Break
12:15 LUNCHEON PRESENTATION I: Optimized Protein and Virus Production in the Expi Expression Systems
Jonathan Zmuda, PhD, Cell Bio R&D, Thermo Fisher
12:45 Luncheon Presentation II (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
1:15 Session Break
Aqua Salon
1:45 PLENARY KEYNOTE PANEL
The PepTalk Plenary Keynote Panel convenes a group of leading scientists working across novel therapeutic modalities and R&D technologies to explore the many challenges associated with discovering, developing, and advancing today’s
novel biotherapeutics. The Panel, via a highly interactive format, encourages discussion among both the panelists and the audience members. Please come prepared with your questions and ideas for this spirited discussion.
- Advances and challenges in expression and production for novel modalities
- Implementing next-generation informatics: data collection, standardization, analysis, ML/AI, and considerations for IP landscape and protection
- Implementing R&D and production capacity for gene and cell therapies – where are we heading?
- Modality-specific challenges: multi-specifics for cancer, improving the ADC therapeutic window, improved safety and pharmacology, novel delivery/targeting
- Preclinical and clinical development of drug combinations with focus in IO: How do we select the right combination dose so we can accelerate clinical development?
Moderator:
Mohammad Tabrizi, PhD, Senior Director, Pharmacology, Ascendis Pharma A/S
PANELISTS
Edward Kraft, PhD, Senior Scientific Manager, Biomolecular Resources, Genentech
Ilya Shestopalov, PhD, Associate Director, Cell Analytics, bluebird bio
David E. Szymkowski, PhD, Vice President, Cell Biology, Xencor, Inc.
Alayna George Thompson, PhD, Senior Scientist I, Drug Discovery Science & Technology, AbbVie
3:05 Refreshment Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
4:00 Chairperson’s Remarks
Johannes Buyel, Dr. rer. nat., Dr-Ing, Head, Bioprocess Engineering, Fraunhofer Institute for Molecular Biology and Applied Ecology IME
4:05 Statistical Design for Effector Function Engineering of Hexameric Fc Domains
Shirley Peters, PhD, Principal Scientist, UCB
We used antibody engineering and recombinant expression to produce Fc with controlled hexa-valency, “Fc-multimer”. Fc-multimer may have clinical potential as a recombinant protein replacement of intravenous immunoglobulin (IVIG)
for the treatment of auto-inflammatory immune disorders. Fc-multimer engages with FcgR’s on multiple cell types and was engineered to maximize potency whilst minimizing potential safety concerns. We used a statistical design approach
(‘Design of Experiments’) to explore the two effector function extremes defined by IgG1 and IgG4.
4:35 Which Sugar Do You Want: Tailormade Glycosylation of Therapeutic Proteins
Bjørn Voldborg, MSc, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark
5:05 High-Level Expression of Thermostable Human Cannabinoid Receptor CB2 in Expi293 Cells for Biophysical Studies
Alexei Yeliseev, PhD, Staff Scientist, Group Leader, LMBB, National Institute on Alcohol Abuse and Alcoholism, National Institute of Health
G protein-coupled receptors (GPCR) comprise a large class of integral membrane proteins important for pharmaceutical drug development. We expressed a full-length cannabinoid receptor CB2 in suspension expi293 HEK cells and purified and solubilized
it in a Façade-TEG detergent. The receptor was homogenous and did not lose its functional activity even after several hours of exposure to elevated temperature. We propose that post-translational modifications of CB2 are essential
for its structural stability.
5:35 Two Cytoplasmic Ubiquitin E3 Ligases and an ER Protease Mediate ER-Associated Degradation of Unfolded Antibody Heavy Chains
Danming Tang, PhD, Technical Development Scientist, Cell Culture, Genentech
Accumulation of unfolded antibody chains in the ER triggers ER stress that may lead to reduced productivity in therapeutic antibody manufacturing processes. We identified UBR4 and UBR5 as ubiquitin E3 ligases involved in HC ER-associated degradation,
while an ER protease PDIA3 cleaves ubiquitinated-HC molecules to accelerate HC dislocation. Proteins characterized in this proteolysis/proteasome-dependent pathway of unfolded antibody HC degradation may be novel targets for CHO cell engineering.
6:05 - 7:00 Networking Reception in the Exhibit Hall with
Poster Viewing (
Sapphire Ballroom)
7:00 Close of Recombinant Protein Expression and Production Conference