| Monday 1 July 2013 |
9.30-10.00 am | Registration and coffee |
Seminar Room A | Registration will take place in the Entrance Hall – you will then be escorted to Seminar Room A for coffee and the day’s proceedings |
10.00-10.15 am | Welcome and introduction |
| Opher Gileadi |
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Session 1 | SOLUBLE/ SECRETED proteins |
10.15-10.35 am 10.35-11.05 am 11.05-11.30 am | Advances in protein production pipelines Chair: Louise Bird Rick Davies: Moving from the single crystal to reproducible crystallisation for multiple ligand cocrystal structures James Love: Eukaryotic protein expression at the New York Structural Genomics Research Consortium Bernadette Mouzon: Generic optimisation using automated small-scale purification |
11.30-12.00 noon | Discussion: Pipelines Chair: James Love |
12.00-1.00 pm | Lunch Lunch will be in the Founders Room |
1.00-1.30 pm 1.30-2.00 pm | Multi-parallel expression strategies; indicators of protein stability/ productivity Chair: Darren Hart Darren Hart: Overview of methods; challenges in library screening in eukaryotic cells Mike Dyson: Screening for functional antibodies |
2.00-2.25 pm 2.25-2.50 pm 2.50-3.15 pm | Expression in E. coli and yeast Chair: Tamar Unger Louise Bird: Using the pOPIN suite of vectors to optimise expression of recombinant proteins Tamar Unger: Protein labelling with FlAsH to monitor protein solubility Barbara Borgonovo: Expression and purification of recombinant human Sonic Hedgehog: E.coli versus mammalian system |
3.15-3.35 pm | Discussion Chair: Nick Berrow |
3.35-4.00 pm | Coffee break (Seminar Room A) |
4.00-4.25 pm 4.25-4.50 pm 4.50-5.15 pm 5.15-5.40 pm | Baculovirus and BacMam expression technologies Chair: Nicola Burgess-Brown Linda King: High titre BacMaM vectors Pravin Mahajan: Baculovirus/insect cell pipeline and BacMam expression Arnaud Poterszman: Tools for expression multi-protein complexes using the baculovirus expression system Jon Elkins: Tags for Kinase purification |
5.40-6.10 pm | Discussion: What limits protein production in baculo? Chair: Nicola Burgess-Brown |
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End of Day 1 |
DAY 2 | Tuesday 2 July 2013 |
Lecture Theatre 5 | The day’s proceedings will take place in Lecture Theatre 5 |
9.00-9.20 am 9.20-9.40 am 9.40-10.00 am | Mammalian expression technologies Chair: René Assenberg David Fisher: Development of a CHO based transient transfection system and its application to a group of secreted proteins Malin Bäckström: The third von Willebrand factor D domain (D3) of intestinal MUC2 mucin forms disulfide bonded trimers and non-covalent hexamers Neesha Dedi: Small-Scale Mammalian Expression |
10.00-10.20 am | Coffee break (Outside Lecture Theatre 5) |
10.20-10.45 am 10.45-11.10 am | Chair: René Assenberg David Hacker: Recombinant protein production from stable Chinese hamster ovary cell lines and pools generated by Piggybac transposon-mediated gene delivery René Assenberg: Transient expression of secreted proteins: HEKm expi293, drosophila; Co-expression systems |
11.10-11.30 am | Discussion: Tips for mammalian expression Chair: David Hacker Including: Economical suspension cultures and transient protein production in CHO lec-minus cells |
11.30-11.50 am 11.50-12.10 pm | Protein engineering, purification and quality control Chair: Paul Carter Adelheid Löhle: Expression, purification and crystallisation of Glucokinase regulatory protein (GKRP) Rod Chalk: Mass spec as part of a high-throughput pipeline |
12.10-12.40 pm | Discussion Chair: Opher Gileadi - Post-translational modification – how to detect and control as required
- Useful co-expression partners/ engineered host strains
- Screening for protein stabilisation: in vivo, in vitro
Rick Davies: Sharing reagents between academia and pharma |
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12:40-1.40 pm | Lunch Lunch will be in the Founders Room |
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Session 2 | MEMBRANE PROTEINS (IMPs) |
1.40-2.10 pm 2.10-2.40 pm 2.40-3.00 pm | Saccharomyces and Pichia pastoris Chair: Radu Aricescu David Drew: Expression of transporters Zarir Unvala: Yeast systems: Pichia vs Saccharomyces (membrane and intracellular proteins) Linda Roth: Expression and purification of Nav1.7 for Antibody Generation |
3.00-3.30 pm | Coffee break (Outside Lecture Theatre 5) |
3.30-3.50 pm 3.50-4.10 pm | Baculovirus/insect cells Chair: Nicola Burgess-Brown Adelheid Löhle: Issues with unsatisfactory purification of GPCRs from insect cells Yin Dong: Optimisation of expression of human IMPs in baculovirus |
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4.10-4.40 pm 4.40-5.20 pm 5.20-5.40 pm | Mammalian systems Chair: David Drew Chris Tate: Expression of GPCRs and transporters using the baculovirus expression system; Expression of serotonin transporter in tet-inducible mammalian expression system T-Rex Radu Aricescu: Transient expression of pentameric ligand-gated ion channels in mammalian cells Paul Carter: Expression and purification of human Phospholipase A2 receptor 1 (PLA2R1) using transient transfection |
5:40-6.15 pm | Discussion: Expression systems for IMPs – what works Chair: Chris Tate |
DAY 3 Seminar Room A | End of Day 2 Wednesday 3 July 2013 |
9.00-9.20 am 9.20-9.40 am | Screening for membrane protein purification conditions, detergents and additives Chair: Simon Newstead Andrew Quigley: Optimising conditions for IMP purification James Love: High throughput protein production and receptor:ligand de-orphaning at the New York Structural Genomics Research Consortium |
9.40-10.10 am 10.10-10.40 am | Mutagenesis strategies for stabilising IMPs Chair: James Love Andreas Plueckthun: Directed molecular evolution of GPCRs Chris Tate: Experiences on the thermostabilisation of three GPCRs for structural studies |
10.40-11.00 am | Coffee break (Seminar Room A) |
11.00-11.40 am | Improving production, stability and quality of IMPs Chair: Liz Carpenter - How to deal with the detergent concentration question
- Which detergents work best for crystallisation for vapour diffusion and LCP?
- Post-translation modification of IMPs – detecting and removing
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11.40-12.00 pm | Conclusions Liz Carpenter |
End of workshop Lunch will be provided at 12 noon in the Founders Room for those who have indicated that they will stay for a networking lunch |