Microbial expression systems offer significant advantages over other hosts by providing faster development times, greater yields, lower production costs, particularly in E. coli which is one of the most widely used hosts for protein production.
However, limitations around glycosylation and central metabolic pathways poses significant challenges.
Cambridge Healthtech Institute’s Microbial Production conference covers the latest developments in microbial expression and production – from host strain development to metabolic engineering, assembly to scale-up, downstream
processing to potential aggregation – with particular focus on the role of E. coli for biotherapeutics, novel products and other industrial applications.
THURSDAY, JANUARY 11
7:45 am Registration and Morning Coffee
8:15 Chairperson’s Opening Remarks
Georg Klima, Dipl. Ing., Executive Director, Process Science, Boehringer Ingelheim RCV GmbH & Co. KG
8:20 Recoded E. coli for Incorporation of Non-Natural Amino Acids and Also for the Purpose of Genetic Isolation
Farren Isaacs, Ph.D., Assistant Professor, Molecular Cellular and Development Biology,
Genetically modified organisms (GMOs) are increasingly used in research and industrial systems to produce high-value pharmaceuticals, fuels, and chemicals. Here, we describe the construction of a series of genomically recoded organisms (GROs), 11
whose growth is restricted by the expression of multiple essential genes that depend on exogenously supplied synthetic amino acids (sAAs).
9:00 Pfizer E. coli Expression Platform Part I: Development and Testing of an Integrated Cloning and Expression System
Kevin Rust, Ph.D., Principal Scientist, Bioprocess R&D, BioTherapeutics Pharmaceutical Sciences, Pfizer, Inc.
9:30 Pfizer E. coli Expression Platform Part II: Application of the Platform to Test Reliably and Rapidly Achieve Increased Protein Yields
Marie Caparon, Ph.D., Associate Research Fellow, Bioprocess R&D, BioTherapeutics Pharmaceutical Sciences, Pfizer, Inc.
The presentation describes the application of an E. coli expression platform to reliably and rapidly achieve increased protein yields. This enables the development of new production processes that are commercially
viable from both a cost of goods and capacity perspective. Case studies will be presented in which the original process presented significant challenges with respect to the cost of goods and the production capacity.
10:00 Coffee Break in the Exhibit Hall with Poster Viewing
11:00 Computational Tools for Improving Microbial Membrane Protein Expression
Bil Clemons, Ph.D., Professor of Biochemistry, Caltech
11:30 Microbial Secretion by ESETEC®: A Cost-Efficient Alternative to Mammalian Cells for Non-Glycosylated Proteins
Sebastian Schuck, Ph.D., Head, Business Development, Wacker Biotech GmbH
The microbial secretion technology ESETEC® offers a cost- and time-efficient alternative for the production of biologics. With straightforward strain and process development, ESETEC
® combines benefits of microbial and mammalian systems. Applying cutting-edge process simulation tools, we demonstrated that Cost-of-Goods for ESETEC® manufacturing
is up to 3 times lower than CHO.
12:00 pm Session Break
12:15 Luncheon Presentation: cGMP Biologics Production Using Corynex
®: A Highly-Productive Gram-Positive Microbial Protein Secretion System
Yoshimi Kikuchi, Ph.D., Executive Professional and Group Manager, Recombinant Protein Research Group, Material Development & Application Labs, Ajinomoto Co., Inc.
Corynex® is Ajinomoto’s highly-productive protein expression system based on the fast-growing gram-positive bacteria C. glutamicum. The powerful and easy-to-handle platform
can secrete correctly-folded proteins directly into the media with high purity and no endotoxins. These advantages allow users to avoid many manufacturing/quality pitfalls and ultimately improve profitability and timelines. We recently
demonstrated successful 1000L cGMP biologics production using Corynex®.
1:15 Ice Cream Break in the Exhibit Hall with Poster Viewing
2:30 Chairperson’s Remarks
Christoph Reese, Ph.D., Director, Microbial Fermentation, Roche Diagnostics GmbH
2:35 E. coli Periplasmic Expression of Antibody Fab Fragments
Mark Ellis, Principal Scientist, Protein Expression and Purification, UCB Pharma
Engineered variants of wild type E. coli strains have been developed which significantly improved periplasmic Fab expression yields. Furthermore, co-expression of E. coli host proteins, combined with engineered strains and fermentation process refinements have enabled Fab yields over 5g/L. These increases have been achieved without compromising cell viability or the quality of the Fab produced.
3:05 Approaches and High Through-Put Tools to Solve Strain and Process Development Challenges for Recombinant Proteins Expressed in E. coli
Nigel Shipston, Ph.D., Director of Program Design, FUJIFILM Diosynth Biotechnologies
3:35 Refreshment Break in the Exhibit Hall with Poster Viewing
4:15 High-Yield, Mineral Medium, Antibiotics Free E. coli Expression System
Christoph Reese, Ph.D., Director, Microbial Fermentation, Roche Diagnostics
The development of a mineral medium based, antibiotics free E. coli expression system will be presented. In a case study, we show the suitability of the expression system for the manufacturing of a recombinant
enzyme in industrial scale in high yield. The system may also be utilized to express otherwise insoluble proteins. A comparison to antibiotics-based selection systems and hydrolysates-based growth media will be discussed.
4:45 Holistic Process Development Strategies for Microbial Expression
Georg Klima, Dipl. Ing., Executive Director, Process Science, Boehringer Ingelheim
RCV GmbH & Co. KG
Novel biotherapeutic formats pose unique development challenges. Strategies for successful development need to holistically consider all aspects of biopharmaceutical processes such as expression strategies, novel unit operations, automated
high-throughput process development, as well as scale up and transfer from bench to large-scale manufacturing. We present our holistic approach based on a HTPD toolbox to lever the complexity of manufacturing development for non-platform
biotherapeutics. Integration of the whole process is also discussed.
5:15 High-Throughput Automated Protein Folding and Quantitative Assessment
Philip An, Scientist, Biologics, Amgen
Employing E. coli to recombinantly express a gene of interest can enable rapid and robust production; however, the proteins are often located in inclusion bodies, necessitating a folding reaction to obtain
their native state. To address this in a high-throughput, quantitative manner, we developed a dense folding matrix system that employs highly integrated liquid-handling automation and a quantitative assessment system to swiftly identify
effective folding conditions.
5:45 Close of Day
FRIDAY, JANUARY 12
8:00 am Registration
8:00 BuzZ Sessions with Continental Breakfast
Protein therapeutics is a fast-growing global market. As the science improves, so does the complexity of the R&D organization. Ensuring product quality plus speed to market requires insights from stakeholders working across the stages
of protein science R&D. Join experts representing this PepTalk pipeline, peers, and colleagues for an interactive roundtable discussion. Topics include highlights from the week’s presentations, new technologies and strategies,
challenges, and future trends.
* Optimizing expression, yields and scale-up
* Process characterization for microbial systems
* Technology gaps and opportunities
Table Moderator: Adekunle O. Onadipe, Ph.D., Associate Research Fellow, Cell Line Development, Bioprocess R&D, Pfizer
9:00 Chairperson’s Remarks
Adekunle O. Onadipe, Ph.D., Associate Research Fellow, Cell Line Development, Bioprocess R&D, Pfizer
9:05 Microbial Process Development in View of Industry 4.0
Peter Neubauer, Ph.D., Prof. Dr., Chair of Bioprocess Engineering, Department
of Biotechnology, Technische Universität Berlin
In our lab-of-the-future concept we integrate and fully automate all wetlab techniques for the production of competent cells, DNA transformation, optimization of cultivation parameters with the analytical methods for medium components
and downstream operation to analyze product activity and other quality parameters. This allows us to apply a dynamic optimal design of experiments strategy (ODoE) to create digital twins for each target protein process (mechanistic
models) which can be applied for computation-based bioprocess development.
9:35 A S. Cerevisiae Process Characterization for Production of a Therapeutic Recombinant Protein Using a Multivariate Analysis
Juan Aon, Ph.D., Senior Manager, MCCD, RD Biopharm, R&D, GSK
10:05 Microbial Expression and Production of Pasylated Proteins and Peptides: Biobetters with Extended Half-Life and Enhanced Action
Uli Binder, Ph.D., CTO, XL-Protein
PASylation comprises the genetic fusion of biologics with a natively disordered biosynthetic polymer made of Pro, Ala and/or Ser (PAS). Such PAS sequences are highly soluble in physiological solution and stably adopt random coil conformation
with an expanded hydrodynamic volume which leads to retarded kidney filtration and drastically prolonged pharmacokinetics in vivo. PASfusion proteins often show improved stability, and they can be
produced in a single step in microbial hosts, thus avoiding costly chemical modification steps.
10:35 Coffee Break with a Poster Pavilion
PepTalk is proud to support and recognize the protein scientists of tomorrow during the Poster Pavilion. This time has been set aside to view the Student Fellowship posters and interact with presenters one on one. This opportunity gives
job seekers the chance to share their expertise with future/potential employers or develop contacts to further their research.
11:15 Towards a New Generation of Glycoengineered Pneumococcal Bioconjugate Vaccines
Christian Harding, Ph.D., CSO, VaxNewMo
Glyco-conjugate vaccines, consisting of a polysaccharide attached to a carrier protein, are excellent immunogens manufactured using labor-intensive chemical crosslinking steps. As an innovative alternative, VaxNewMo utilizes a glycoengineering
strategy to generate “bioconjugates” in Escherichia coli. Key to this process is a conjugating enzyme, which attaches a polysaccharide to a protein.
11:45 Expression of Complex Proteins in E. coli
Dorothea Reilly, Ph.D., Principle Scientist and Associate Director, Early Stage Cell Culture, Genentech
Genentech produces recombinant proteins for therapeutic use employing both CHO cells and E. coli. We have developed E. coli processes that are optimized for the secretion
of recombinant proteins into the periplasmic space where folding and assembly can occur. This talk will describe how these approaches have enabled the production of complex protein formats.
12:15 pm Conference Wrap-Up
Adekunle O. Onadipe, Ph.D., Associate Research Fellow, Cell Line
Development, Bioprocess R&D, Pfizer
12:45 Close of Conference