The Bioprocessing Summit
2013 Archived Content

Short Course 5: E. coli Innovations – Dinner Short Course

Tuesday, August 20, 2013 | 6:00 to 9:00pm

 

6:00    Engineered E. coli Host Strains for Periplasmic Protein Production

David P. HumphreysDavid P. Humphreys, Ph.D., Director, Antibody Biology, UCB-New Medicines

E. coli remains an attractive expression host due to its potential for high biomass, large scale and short cycle production processes. In particular, small antibody fragments are well suited to expression in E. coli. This presentation will cover various aspects of strain engineering illustrated with examples from our own experience and from the literature. We have engineered variants of wild type E. coli strains in order to significantly improve periplasmic expression yields. In combination with plasmids co-expressing E. coli host proteins and process refinements we have achieved purified Fab’ yields in the 2 to 5g/l range with a number of clinically relevant Fab’s. These technological improvements have also enabled the expression of a variety of ‘difficult’ proteins at several hundred mg/l and hence have the potential for a broad impact on both the production of novel therapeutic formats and supply of research reagents.

6:45    Dinner Break 

7:15    A Platform Manufacturing Process for Fabs Produced with E. coli Secretion Technology  

Susanne DilsenSusanne Dilsen, Ph.D., Head, Production, Wacker Biotech GmbH

Antibody fragments are a current trend in the biopharmaceutical industry as they are less complex than antibodies and yet still retain high target specificity. Using Wacker’s E. coli-based secretion technology ESETEC® for the expression of Fab antibody fragments, an efficient and scalable platform manufacturing process was developed at 300 L fermentation scale. Selected case studies will be shown, along with the advantages of the Fab-E.coli approach versus the classical IgG-CHO approach regarding speed-to-clinic and cost of goods.

8:00    Production of Homogeneous Best-in-Class Antibody Drug Conjugates by Cell-Free Protein Expression
in E. coli
  

Trevor J. HallamTrevor J. Hallam, Ph.D., Chief Scientific Officer, Sutro Biopharma, Inc.

Sutro has developed a robust, scalable and controllable cell–free protein synthesis platform for the efficient production of homogeneous therapeutic proteins, including full length IgGs and ADCs, bispecific antibodies, cystine-knot peptides and proteins. Many variants can be expressed in hours and rapidly assessed for function. Within days, production of chosen variants can be scaled using the same platform to generate material for GLP-tox studies and clinical studies. The platform brings speed to molecular design, process development and manufacture and could enable pre-clinical timelines to be reduced by 18-24 months from concept to clinic.


SPEAKER BIOGRAPHIES

David P. Humphreys, Ph.D., Director, Antibody Biology, UCB-New Medicines

David Humphreys graduated with a degree in molecular biology from the University of Durham (UK) in 1992 before completing a PhD at the University of Birmingham (UK) in 1995. His PhD investigated the folding and augmented expression of heterologous proteins in the E. coli periplasm, using protein disulphide isomerase as a chaperone. David joined UCB (formerly Celltech) in 1995 where he first applied his scientific skills to many aspects of the engineering of antibody constructs and their expression in various hosts. Research interests have included; Fab and antibody engineering, E. coli strain engineering, protein secretion and folding and improved PEGylation strategies. David now manages a team of researchers working on various aspects of antibody engineering and microbial and CHO expression. His work has resulted in multiple peer reviewed publications, reviews and granted patents.

Trevor J. Hallam, Ph.D., Chief Scientific Officer, Sutro Biopharma, Inc.

Dr. Hallam brings more than 25 years of experience in drug discovery and development toSutro. Most recently, he was executive vice president of research & development and executive officer at Palatin Technologies where he was responsible for discovery and clinical development and led the development of several INDs for drug candidates in a variety of therapeutic areas. Prior to joining Sutro, Dr. Hallam held several senior management positions at AstraZeneca, most recently vice president, biologics (respiratory and inflammatory diseases), vice president, U.S. medical affairs, and vice president, preclinical research and development, respiratory and inflammation, Sweden. He began his pharmaceutical career at Smith Kline & French Research, U.K., followed by senior management positions at Glaxo Group Research, U.K., Roche Research Centre, U.K., and Rhone Poulenc Rorer, U.S. Dr. Hallam conducted his post-doctoral training in the laboratory of Timothy J. Rink, M.D., Sc.D., at the Physiological Laboratory, University of Cambridge after receiving his Ph.D. in biochemistry from King's College, University of London.

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