The course is designed primarily for structural biologists who have prokaryotic expression experience and would like to use the mammalian expression system for more challenging targets. The goals of the course are to allow participants to have hands-on experience of mammalian cell culture, small and large scale transient gene expression including the use of automated systems, protein purification and crystallisation. The course will be structured so as to interleave lectures and practical sessions. The participants will have the opportunity to closely interact with the tutors and to directly discuss their research projects with the experts in small workgroups.
Participants will be asked to submit their CV and a letter of motivation. The course will be limited to 12 participants from European structural biology laboratories.
1. Mammalian expression platform overview
2. Mammalian cell lines for protein expression and glycosylation control
3. Mammalian expression constructs design
4. Small scale expression screen with Western Blot
5. Large scale expression and protein purification
6. Protein crystallisation
7. Protein structure case studies
Radu Aricescu (Oxford University, UK), Yuguang Zhao (Oxford University, UK), Joop van den Heuvel (Helmholtz-Zentrum für Infektionsforschung, DE), Veronica Chang (Oxford University, UK), Ben Bishop (Oxford University, UK), Karl Harlos (Oxford University, UK), Tom Walter (Oxford University, UK), Christian Siebold (Oxford University, UK), Joerg Standfuss (Paul Scherrer Institut, CH)