The 8th 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. This conference
poses the question: ‘What is the best expression system to use to fulfill a project’s goals?,’ while staying within budget and adhering to project timelines. Experts will share case studies and disclose data, while divulging
details of these systems’ underlying mechanisms. Comparing and contrasting systems will also be featured to increase understanding in the quest for greater productivity.
SC12: Transient Protein Production in Mammalian Cells
Richard Altman, MS, Scientist, Protein Technologies, Amgen
Henry C. Chiou, PhD, Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific
*Separate registration required.
WEDNESDAY, MAY 2
7:30 am Registration(Commonwealth Hall) and Morning Coffee (Harbor Level)
8:30 Chairperson’s Remarks
Jesse Rinehart, PhD, Associate Professor, Cellular & Molecular Physiology, Systems Biology Institute, Yale University School of Medicine
Jorg Thommes, PhD, Senior Vice President, Pharmaceutical Sciences & Technology, Visterra, Inc.
A robust manufacturing platform for recombinant protein and antibody manufacturing has emerged over the past decade, which has substantially increased productivity and operational reliability. Thus, manufacturing has matured from a “necessary
evil” to a true value driver in biopharmaceutical operations. In this presentation, we will discuss the current state of protein drug manufacturing but also review new frontiers in this field, e.g., further industrialization of antibody
manufacturing, accelerated manufacturing for early clinical testing, and manufacturing for new therapeutic modalities.
9:10 Implementation of New Strategies During Early Development of Innovative Therapeutic Molecules
Nicola Beaucamp, PhD, Head, Process Research, Roche Innovation Center Munich, Pharma Research and Early Development,
Roche Diagnostics GmbH
More than 80% of large molecules developed in Roche pRED are novel and complex antibody formats. To minimize timelines and deliver requested quantity and quality, many innovative approaches with respect to throughput, automation and cell culture
techniques were taken. These techniques were implemented to support delivery of differentiated molecules based on Roche’s strategy on engineering technologies to patients. Technical challenges faced and how they were successfully solved
will be presented by case studies.
9:40 Revolution or Evolution – Optimizing Protein Expression Platforms to Support Preclinical Research at AstraZenca
Robert Roth, PhD, Associate Principal Scientist, Innovative Medicines, Discovery Sciences, AstraZeneca
An overview of how AstraZeneca have refined and developed our protein expression platforms to supply reagents to support research across a diverse portfolio. Using specific examples to illustrate how challenges have been resolved through internal
development efforts, collaborations with academic partners and technology providers. How do we approach a future where targets are increasingly complex and require bespoke solutions to identify and express proteins that are physiologically
10:10 Coffee Break in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
10:55 Isolation of CHO Master Cell Lines and Targeted Cell Line Engineering using Recombination Mediated Cassette Exchange (RMCE) and Homology-Directed Repair (HDR)
Joop van den Heuvel, PhD, Research Group Leader, Recombinant Protein Expression, Helmholtz Centre for Infection Research
Chinese Hamster ovary (CHO) cells are one of the major providers of recombinant protein for biopharmaceutical industry and biomedical research. Long tradition in laboratory handling generated a wealth of experience in engineering CHO cell lines with a variety of genetic tools to achieve efficient production of recombinant proteins. Targeted manipulation of CHO cell lines based on recombinase-mediated cassette exchange (RMCE) and CRISPR-Cas-induced homology-directed repair (HDR) to generate high-expression CHO cell lines will be presented. High-expressing master cell lines have been isolated, which carry tagged expression cassettes. These cell lines are transferred with the help of a “selection trap” into producer cell lines by the exchange reaction. The efficiency of the isolation of clonal producer cell lines and quality of the protein production for challenging targets will be discussed.
11:25 A Versatile Platform for Biologics Research Protein Reagent Generation
Bernd Voedisch, PhD, Lab Head and Investigator III, NIBR Biologics Center, Novartis Institutes for Biomedical Research
During the early phases of Biologics research projects, it is necessary to produce a plethora of high quality protein reagents and recombinant cell lines in order to enable generation, screening, and in-depth characterization of therapeutic
antibody candidates. The presentation will highlight various challenges in this context, and how these challenges were overcome by applying cutting-edge technologies (e.g., cell line engineering using CRISPR/Cas9 or transposases) in order
to lay the foundation for successful Biologics discovery.
11:55 MultiBac: From Protein Complex Structures to Synthetic Viral Nanosystems
Imre Berger, PhD, Professor and Wellcome Trust Senior Investigator, Biochemistry, Biomedical Sciences and BrisSynBio Centre,
University of Bristol
The MultiBac BEVS was conceived as a user-friendly tool-kit for producing multiprotein complexes for structural biology, and enabled structure determination of many molecular machines, including previously inaccessible high-value drug targets.
More recently, MultiBac developments focus on customized baculoviral genomes tailored for specific applications, such as synthesizing artificial proteins by genetic code expansion, and DNA delivery in mammalian cells and tissues for CRISPR/Cas9-mediated
gene editing. Some of these developments will be presented.
12:25 pm Accelerating Gene to Phase 1 Studies: Ultra-Rapid Multi-Dimension Screening Tools and Optimized Scale up Strategy to Deliver the Drug Substance from E coli
Prabuddha Kundu, PhD, Executive Director, CEO, Premas Biotech Pvt Ltd
With modern drug discovery, shortening to time to clinics is of utmost importance and a significant challenge. Premas has generated ultra-rapid methods, screening tools and scale up strategy to deliver a scalable process, validated drug substance
for enabling a Phase 1 trial. Screens were established, optimized and operation excellence was implemented to determined the optimum conditions. This was accomplished within 22 weeks from the gene synthesis to deliver of drug substance.
12:55 Luncheon Presentation I: New Tools for Screening & Harvesting Solutions for CHO & HEK293 Cells, for both Transient and Stable Cell
Daniel Korostyshevsky, Tri-State Technical Sales Representative, 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.
1:25 Luncheon Presentation II: Accelerating Lead Magnetic Bead Purification Technology
Ray Low, PhD, Scientist, Amgen Inc.
High throughput purification is a critical step in antibody engineering and lead identification. Magnetic based technologies allow single step purification of targets by adding beads directly into bioreactors. This eliminates sample clearance
through centrifugation - filtration and loading voluminous feedstocks to columns. Here, Amgen presents one of its latest innovations in the field of antibody screening and drug discovery using several parallel HTP platforms. Some of these
systems are completed and licensed to GenScript.
1:55 Session Break
2:10 Chairperson’s Remarks
Bernd Voedisch, PhD, Lab Head and Investigator III, NIBR Biologics Center, Novartis Institutes for Biomedical Research
2:15 The Application of Next-Generation Sequencing to Understand CHO Cell Biology
Colin Clarke, PhD, Principal Investigator, National Institute for Bioprocessing Research and Training (NIBRT)
The publication of genome sequences for CHO cell lines has facilitated study of these cell factories with unprecedented resolution. Our understanding of the CHO cell transcriptome, in particular, has rapidly advanced through the application
of next-generation sequencing (NGS) technology to characterize RNA expression (RNA-Seq). In this talk, I will present examples of how RNASeq is being utilised to enhance the production of biopharmaceuticals in CHO cells.
2:45 Application of 13C Flux Analysis to Identify High-Productivity CHO Metabolic Phenotypes
Jamey Young, PhD, Associate Professor, Chemical and Biomolecular Engineering, Vanderbilt University
Identifying metabolic phenotypes that promote high expression is a major goal of the biotech industry. We conducted a series of 13C flux analysis studies to examine the metabolic response to IgG expression during early stationary
phase of CHO cell cultures. Lactate consumption and citric acid cycle fluxes were most strongly associated with specific IgG productivity. These studies indicate that enhanced oxidative metabolism is a characteristic of high-producing
CHO cell lines.
3:15 Implications of Choosing the Right Cell Engineering Technology on Early Stage Process Development
James Brady, PhD, Vice President, Technical Applications and Customer Support, MaxCyte
Increasing productivity & maximizing data relevance during biotherapeutic development is possible when manufacturing the host cell early to produce sufficient quantities of candidates for extensive characterization and progression of promising
candidates. Employing proper cell engineering, such as MaxCyte’s high-performance cell engineering technology, drives rapid, scalable production of proteins in the manufacturing cell background while maintaining flexibility in downstream
culture processes. This presentation discusses how a technology positively impacts productivity and fast-tracks biotherapeutic development.
3:45 Refreshment Break in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
4:45 Problem-Solving Breakout Discussions (Commonwealth Hall)
Non-Standard Amino Acid Incorporation in Recombinant Proteins
Moderator: Jesse Rinehart, PhD, Associate Professor, Cellular & Molecular Physiology, Systems Biology Institute, Yale University School of Medicine
- The unique challenges of incorporating non-standard amino acids
- The choice of cellular host
- Incorporation strategy
- Basic set-up/trouble-shooting
- Mass spectrometry/proteomics of non-standard amino acid containing proteins
5:45 Networking Reception in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
7:00 pm End of Day
THURSDAY, MAY 3
8:00 am Registration(Commonwealth Hall) and Morning Coffee (Harbor Level)
8:30 Chairperson’s Remarks
Nicola Beaucamp, PhD, Head, Process Research, Roche Innovation Center Munich, Pharma Research and Early Development, Roche Diagnostics GmbH
8:35 Massively Parallel Human Systems Biology in Synthetic Organisms
Jesse Rinehart, PhD, Associate Professor, Cellular & Molecular Physiology, Systems Biology Institute, Yale
University School of Medicine
Protein phosphorylation encompasses a central cellular language that determines every facet of normal cellular biology. We have recently created a technology that enables site-specific incorporation of phosphoserine into proteins by expanding
the genetic code of Escherichia coli. I will describe our new capability to synthesize and observe phosphoproteome-scale libraries of human phosphoproteins that enable answers to systems level
questions regarding the functional role of all human phosphorylation events.
9:05 Evolving and Engineering E. coli Recombinant Protein Production Strains
Jan-Willem de Gier, PhD, Professor, Biochemistry and Biophysics, Stockholm University
My laboratory has used both evolutionary and engineering approaches to create E. coli strains with improved properties for recombinant protein production. The evolutionary approaches will be illustrated
by the isolation of BL21(DE3)-derived membrane protein production strains. The engineering approaches will be illustrated by the construction of an E. coli strain background with tighter regulation
of rhamnose-induced protein production.
9:35 Myceliophthora Thermophila, “C1” a Recombinant Protein Production Platform
Mark Emalfarb, CEO, Dyadic International, Inc.
Dyadic has developed a novel eukaryotic gene expression platform with improved properties to speed up the development and lower the manufacturing cost of recombinant proteins. Intensive glycoengineering work is being done to develop cell
lines to produce proteins at high yields, using low cost defined media with defined human glycoforms.
10:05 Coffee Break in the Exhibit Hall with Poster Viewing (Commonwealth Hall)
11:05 RAF1 Kinase: Everything but the Kitchen Sink
William Gillette, PhD, Principal Scientist, Deputy Director, Protein Expression Laboratory, Leidos Biomedical
RAF1 kinase is a critical protein in the regulation of cell proliferation in normal cells and is the focus of research due to its role in cancer. Obtaining highly purified, active RAF1 in quantities sufficient for HTP assays or structural
biological research has not been reported in the literature. I will present a summary of our efforts to express and purify RAF1, and the impact of expression system, cell line, temperature, and length of expression.
11:35 Development-Oriented Biologics Discovery
Bingyuan Wu, PhD, Senior Research Scientist, Janssen Pharmaceutical Companies of Johnson and Johnson
While the biologics market is expanding rapidly with novel and effective therapeutics, the challenges in discovery are steadily increasing. Discovery must be nimble and seamless with development. Recent examples will be discussed that
demonstrate Janssen’s robust and efficient expression systems to support and profile innovative protein therapeutics that make up the early portfolio.
12:05 pm Optimisation of Transient Expression Platform to Increase Titre and Throughput
David Humphreys, PhD, Director and Head, Protein Sciences, UCB Pharma
12:35 End of Optimizing Protein Expression