Transcript Synthelis structural biology presentation february 2016

Bringing structural biology
services through collaboration
1/ Collaboration key players
Synthelis and ESRF
SYNTHELIS / www.synthelis.com
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Synthelis, a dynamic company
Transforming academic expertise into relevant services
 Spin-off company from University Joseph Fourier
Grenoble - Incorporated in January 2011
 Bringing service offer to industries : development,
production & characterization of difficult to express
proteins (membrane & cytotoxic proteins)
Proprietary technology and
know-how in cell-free protein
expression
SYNTHELIS / www.synthelis.com
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Recognized expertise
Already validated expertise with well-established customers
 33 client portfolio
 93% of protein successfully expressed
 77% with proof of activity
 > 80 proteins successfully produced to date
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Structural biology cluster – Grenoble area
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ESRF in brief
European Synchrotron Radiation Facility (ESRF)
 Established in 1975 with the synchrotron project,
capable of producing highly brilliant hard X-rays.
Services
 Beam time measurements
 Experiment support by in-house experts
 Industrial access
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Simple and effective access to X-ray facilities and expertise
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Specialized laboratory services
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Technology transfer, instrument sales and licensing
World reference for
industrial and academic use
of synchrotron radiation
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2/ Integrated offer
From risk analysis to MAD/SAD structure solving
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Overall process*
10 to 14 months
1 week
5 - 6 months
Se-Met
expression
Risk analysis
Expression
upscale
1 month
Stability and
activity
assessment
(TSA)
1–2 month(s)
Purification
set-up &
optimization
1-2 month(s)
Cristallogenesis
1 week
1 week
Diffraction
test
Data
collection
Biophysics
options
If no crystals
2 weeks
MAD/SAD
structure
solving
If bad
diffraction
Crystal
optimization
1-2 month(s)
* Approximated timelines
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Risk analysis
Service offer
 Sequence analysis and structure prediction tools
 Recommendation on which or how many construct(s)
to be tested
 Reporting and scientific meeting for GO/NO GO decision
Sequence alignement
Timeline
1 week
Disorder prediction (Source: Thesis from E.Poudevigne)
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Se-Met expression & expression upscale
Service offer
 Two genetic construction design and production
 Screening of cell-free expression conditions using Se-Met amino acids
 Pre-purification by affinity
 Western-Blot for identification and Coomassie Blue Staining gel for quantification
 Expression scale up to produce mgs for characterization, purification and
crystallogenesis.
Timeline
5 - 6 months
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Stability and activity assessment (TSA)
Service offer
 For the 2 constructions in different conditions
 Thermal shift assays to assess the activity and the thermo-stability of the proteins
 Reporting and scientific meeting for GO/NO GO decision
Timeline
1 month
TSA measurements
SYNTHELIS / www.synthelis.com
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Purification set-up & optimization
Service offer
 Purification with pre-determined protocol and concentration of sample up to 10mg/ml
if possible
 Concentration and solubility tests
 Reporting
Timeline
1-2 month(s)
(Source: Thèse E.Poudevigne)
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Crystallogenesis
Service offer
 Crystallization set-up
 Image Inspection/annotation
 Screening of 1 protein samples up to 3 concentrations
(one screening consisting of 5-6 crystallization plates
+1 refinement plate).
Preparation of cryogenic solutions for up to best 5 hits
per screen.
 Reporting
Timeline
1-2 month(s)
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Biophysics options
Service offer
 Size and shape of macromolecules
 Stoichiometry of macromolecular assemblies
 Secondary and tertiary structure of macromolecules
 Thermodynamic stability of macromolecules
 Properties kinetic and thermodynamic interactions
Timeline 2-6 weeks
depending on options
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Diffraction test
Service offer
 Test of crystal hits on synchrotron beamline
 Feedback to crystallisation if further refinement
required
Timeline 1 week
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Crystal optimization
Service offer
 Manual production of crystals for data collection with optimized conditions from (5).
Production of cryo-conditions, mounting and freezing crystals.
Maximum duration of three months.
 Manual crystal optimization
 Consumables
 Reporting
Timeline
1 – 2 month(s)
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Structure solving and refinement
Service offer
 Protein structure determination – via molecular
replacement or use of anomalous scattering
 Subsequent refinement of structure
 Reporting to level equivalent to PDB structure deposition
Timeline 2 weeks
for standard resolution
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3/ Why work together ?
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Benefits
 Complementary expert teams
 Proximity between the production site and the structure determination site
 Single point-of-contact to access the whole value chain
 Industry-driven service offer
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Your contact at SYNTHELIS :
Alex Abadie
5 Avenue du Grand Sablon
38700 Grenoble - FRANCE
Phone : +33 4 76 54 95 35
Mobile : +33 6 46 23 12 60
[email protected]
www.synthelis.com
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