Monday, 2 November, 09:00 – 12:00

SC3: CHO Cell Engineering

Co-Instructors:

Anton Bauer, Ph.D., COO, The Antibody Lab

Simon Fischer, Ph.D., Scientist, BP Process Development Germany, Boehringer Ingelheim Pharma GmbH & Co. KG

Zhiwei Song, Ph.D., Principal Scientist, Lead PI for GlycoSing Programme, Bioprocessing Technology Institute, A*STAR

Recombinant protein therapeutics have proven their worth as invaluable pharmaceuticals. Chinese hamster ovary (CHO) cells are the primary choice by industry for the production of these proteins, owing to their capacity for correct folding, assembly and posttranslational modification. The growing demand for therapeutic proteins necessitates the development of new technologies for high quality and productivity in CHO expression systems. This course explores CHO cell engineering strategies to improve and select for the highest producers.

09:00 Welcome and Introductions

09:15 Improved Protein Production Using Euchromatin-Containing BAC Expression Vectors in CHO Cells

Anton Bauer, Ph.D., COO, The Antibody Lab

We apply an active gene environment from BACs, rather than specified genetic elements, in expression vectors used for random integration and achieve high-recombinant protein production in CHO cells. Clonal protein production is achieved quickly, and is directly proportional to integrated vector copy numbers and remains stable without selection pressure. Production is about tenfold increased, and we show examples for model proteins and antibody compared to conventional vectors.

10:00 Making Cell Engineering Using MicroRNAs More User Friendly: A Guide towards Successful MiRNA Applications in Mammalian Production Cell Systems

Simon Fischer, Ph.D., Scientist, BP Process Development Germany, Boehringer Ingelheim Pharma GmbH & Co. KG

Cell engineering of biopharmaceutical production cell factories using non-coding RNAs to enhance culture performance is currently in vogue. MicroRNAs (miRNAs) have recently gained much attention as powerful molecular tools to improve cell phenotypes without adding translational burden to the cell. However, some users are still restrained for giving miRNAs a try in their host cell systems. We introduce practical applications to rapidly identify and exploit impactful miRNAs for cell engineering.

10:45 Coffee Break

11:00 Generation of CHO Glycosylation Mutants for the Production of Recombinant Therapeutics with Enhanced Efficacy

Zhiwei Song, Ph.D., Principal Scientist, Lead PI for GlycoSing Programme, Bioprocessing Technology Institute, A*STAR

A panel of novel CHO glycosylation mutants have been isolated/generated by cytotoxic lectins and the genome editing technologies. These mutants can be used to produce highly sialylated glycoproteins, glycoproteins with mannose-terminated N-glycans and fucose-free antibodies. The DHFR and GS genes in these cells have been inactivated as selection markers and the cells have been adapted to robust growth under serum-free suspension conditions.

11:45 Interactive Q&A with Instructors and Participants

12:00 Close of Short Course

Instructor Bios:

Anton Bauer, Ph.D., COO, The Antibody Lab

After finishing his studies in biochemistry and immunology at the University of Vienna, Anton performed his Ph.D. studies in cell biology and oncology at the Research Institute of Molecular Pathology of Boehringer Ingelheim in Vienna. Upon graduation in 1998 he continued postdoctoral studies in developmental biology and immunology at the German Cancer Research Center in Heidelberg. When he returned to Austria, he focused on translational sciences in biotech companies mainly dealing with antibodies. In 2006 he joined the founding team of the startup company F-star GmbH in Vienna, where he developed the next-generation antibody platform with his team. He then moved to Intercell (Valneva Austria), where he headed the endogenous human antibody platform. Recently he joined the management team of a new endeavor in Vienna, "The Antibody Lab" (www.theAntibodyLab.com). Together with his colleagues he established the use of euchromatin-containing BAC expression vectors for rapid and stable recombinant protein production in CHO cells.

Simon Fischer, Ph.D., Scientist, BP Process Development Germany, Boehringer Ingelheim Pharma GmbH & Co. KG

Simon Fischer works as a scientist at the Department of BP Process Development at Boehringer Ingelheim Pharma, Biberach, Germany and is responsible for implementing novel technologies in Cell Line Development. Simon acquired his Ph.D. in Pharmaceutical Biotechnology at the Institute of Applied Biotechnology Biberach, as a fellow of the International Graduate School in Molecular Medicine of Ulm University. Before, he received the Master of Science (MSc) degree in Biomedical Engineering from the University of Applied Sciences Albstadt-Sigmaringen and the Bachelor of Science (BSc) degree in Pharmaceutical Biotechnology from the University of Applied Sciences Biberach. He also gained relevant industry experience by working with Novartis Pharma AG at Basel, Switzerland, as well as Boehringer Ingelheim Pharma. Furthermore, he was guest researcher at the Donders Institute for Brain Function, Cognition and Behaviour at the Radboud University Nijmegen, The Netherlands. Simon built up the miRNA research platform in mammalian production cell systems at the IAB Biberach, including the development of various cell-based high-content miRNA screening systems.

Zhiwei Song, Ph.D., Principal Scientist, Lead PI for GlycoSing Programme, Bioprocessing Technology Institute, A*STAR

Dr. Zhiwei Song obtained his Ph.D. degree in Biochemistry from the University of Michigan where he worked on protein glycosylation. His postdoc work was done in the Biology Department at MIT where he studied the genetic control of apoptosis in Drosophila. He then worked in Dr. Daniel I.C. Wang’s lab at MIT for a year and a half. In 2002, Zhiwei moved to Singapore. Now he is a Principal Scientist at the Bioprocessing Technology Institute (BTI) of A*STAR. His research has been focused on protein glycosylation, production of recombinant antibodies using CHO cells and apoptotic cell death in cultured mammalian cells.