The 2nd SGC Protein Production Workshop

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Details

A three-day workshop on the challenges and developments in recombinant protein production.

The SGC is hosting a workshop bringing together ~50 scientists from academia and industry to discuss the current state of recombinant protein production. Topics include: expression in bacteria, baculovirus and mammalian cells; high-throughput approaches; optimization of expression and purification; optimizing protein stability; quality control; and integral membrane protein (IMP) production. We plan to devote Monday and Tuesday morning to soluble proteins and general issues; Tuesday afternoon to IMPs, and Wednesday morning to more specialized issues related to IMPs.

The workshop will consist of topical sessions; participants are asked to submit suggestions for short (10 minute) presentations. The discussion leaders of each session will then arrange a sequence of short presentations followed by a 10-15 minute open discussion. We hope this format will promote a frank discussion of the challenges and a critical evaluation of the utility and impact of new and old technologies.

The proposed topical sessions are listed in the Programme tab. To propose a presentation, please follow the instructions through the registration process.

Registration

Participation is free of charge. Buffet lunch and coffee will be provided during the meeting. We are unfortunately not able to cover travel and accommodation for delegates or speakers.

The number of participants is limited to 60, so registration before the deadline of May 15th is required. Participants are encouraged to submit proposal for short presentations, but this is not a condition for attendance. A confirmation of registration will be sent immediately; however, final confirmation of participation will be sent only after the registration deadline.

Abstract submission

The meeting agenda depends on your input.

Please submit proposals for short (10 min) presentations; you can register to present in 1-3 sessions. We will try to accommodate as many presentations as possible within reasonable session lengths. If you propose more than one presentation, please rank in order of preference, in case we cannot accommodate all.

Please submit your proposals by May 15^th.

Programme

Below is the full programme for the day. You can download the full programme in PDF format here

DAY 1

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

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

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

12:40-1.40 pm

Lunch

Lunch will be in the Founders Room

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

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

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

Location

The workshop will be held at the Said Business school in the centre of Oxford, next to the main railway station.

You can find out more about the Said Business School by visiting their website

The location of the Said Business School is shown on a map here

Accommodation

We are unfortunately unable to cover the cost of accommodation in Oxford during the workshop. If you require accommodation, we suggest that you book early as Oxford is a popular destination in summer. The below links may be useful in providing a starting point in your search for accommodation:

Oxford University rooms: http://www.oxfordrooms.co.uk/

Hotels, B&Bs, etc: http://www.visitoxfordandoxfordshire.com/oxford/

Please note that the workshop venue is not close to the SGC offices and laboratories at Headington; the Saïd Business School Park End Street campus is instead next to the railway station (its postcode is OX1 1HP).

Speakers

Speakers list TBC

Please check back for a list of confirmed speakers at this workshop.

Registration is now closed. Please contact protproduction2013@thesgc.org for further enquiries.