Institute for International Research

Description

This course introduces the fundamental concepts needed to establish small to large-scale transient protein production systems using mammalian cells. The class examines in detail the four essential elements of any mammalian transient production system: cell lines, expression vectors, transient transfection and cell culture. The course helps participants understand differences and tradeoffs in producing recombinant proteins in HEK293, CHO, or other mammalian cells. We review expression vector basics, delve into advanced vector optimization and examine scale-up considerations for transient transfection. Specific attention is focused throughout the course on considerations for production of monoclonal antibodies. The most commonly used transfection reagents and transfection methods are examined. Attendees gain an understanding of the equipment needed to establish a transient production facility, methods to monitor culture conditions and how to assess transfection efficiency.

Agenda

• Overview and Introduction to Mammalian Transient Expression Systems
  • rationale for transient protein production
  • comparison to non-mammalian production systems
  • comparison to stable expression systems
• Cell Lines for Transient Production
  • the HEK293 versus CHO decision
  • alternative mammalian cell lines
  • host cell differences in post-translational processing
  • adherent and suspension culture
  • engineered hosts for increased production
• Expression Plasmids
  • expression vector engineering
  • leading edge cloning strategies
  • coding sequence optimization
  • antibody expression vector strategies
  • plasmid production
• Concepts in Large-Scale Transient Transfection
  • transfection reagents and methods
  • transfection optimization
  • transfection analysis and monitoring
• Cell Culture
  • culture medium for transient transfection
  • cell culture analysis and monitoring
  • optimizing culture conditions
  • culture supplements and enhancements for increased production
• Equipment for Transient Protein Production
• Scale Up
• Additional Topics and Cost Considerations

Who Should Attend?

• Scientists needing to quickly produce milligram to gram quantities of recombinant protein in mammalian cells
• Scientists, Managers and Lab Directors establishing core protein production services for drug discovery
• Bioproduction and Process Development Scientists wanting to learn about transient protein production methods
• Protein expression Scientists looking to scale up production

Instructor

Henry C. Chiou, Ph.D., Director, Research and Development, Invitrogen Corporation
Henry Chiou has over 15 years of experience in mammalian expression technology and in nucleic acid delivery systems. He has broad expertise in expression vectors, cloning, cell biology and cell culture. Henry directs Invitrogen's R&D efforts in the development of new transfection reagents and transfection applications. Dr Chiou obtained a bachelor's degree from Yale University in biochemistry and earned a doctorate in Molecular Pharmacology from Harvard University. He then completed a post-doctoral fellowship studying viral expression elements at the University of Pennsylvania. Prior to joining Invitrogen he worked for a number of years in small to midsized biotech companies developing biotherapeutics.