1 / 19

Working Group 1 Planing 1st Semester 2013

Working Group 1 Planing 1st Semester 2013. Planing Overview. General Plan Tasks Description Deliverables : 1st Semester 2013 WG1: Recruiting Groups. Planing Overview. Non- characterized Proteins. Recombinant Proteins. Planing Overview. Non- characterized Proteins.

strom
Télécharger la présentation

Working Group 1 Planing 1st Semester 2013

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. WorkingGroup1 Planing 1st Semester 2013

  2. PlaningOverview • General Plan • TasksDescription • Deliverables: 1st Semester 2013 • WG1: RecruitingGroups

  3. PlaningOverview Non-characterizedProteins RecombinantProteins

  4. PlaningOverview Non-characterizedProteins RecombinantProteins Universidad de Salamanca-CSIC Clones Collection ( 9800)

  5. Clones Collection Clones Collection ( 9800) Clones Collection ( 13500) 13500 clones ( 2013-2014)

  6. RecombinantProteins In situ Proteinexpression ConventionalProteinexpression •Fast ●cDNAavailability ●Clones sequencevalidated ●Yield ? •Slow ●Subcloning ●Experience: E.coli, CHO, Yeast… ●HighYield ( purification ?)

  7. In situ ProteinExpression • IVTT: Rabbits vs HeLA

  8. In situ ProteinExpression • IVTT: Rabbits vs HeLA • OptimizedprotocolsforProteinEnrichement • MS assays ( CICBiogune, UCM, ???) • 25 proteinsexpressedbyHeLa IVTT ( CIC) • 50 additional clones validated ( UCM & CIC) • Training of HeLa IVTT at CIC ( open)

  9. ConventionalProteinExpression PreliminaryCriteria • Cloning & Subcloningstrategies • Selection of expressionsystem: E.coli, CHO, yeast, Pichia,…) • Non-succesfully IVTT expressedproteins • Highdemand of recombinantproteinsbyotherWGs

  10. WorkingFlowbetweenWG1 & WG-MRM • Deliverable: Data, purifiedprotein, IVTTmix+protein, gel band, protocols,…. • Amount of protein: ~ 20 µg / Lab • Number of Labsfor MRM: 3/protein ??? • Updatedlist of unknown/non-characterizedproteins • Number of labsfor MRM: Recruiting

  11. Cloning • No in CIC collection (9800) ( 642 chr16) • No in ASU collection (13500) ( > 642 chr16?) • Cloning & subcloning: Afterfirstyear A fewselectedproteins Conventionalcloning $$$ Othersources $$$$

  12. BudgetScouting • 600 €/ ProteinHeLa IVTT for 200 µg • 250€/full process per protein ( < 20 µg) • Milestone: 300 proteins / 2 years 60 k€ ( personal cost no included)

  13. Nowis time for • Suggestions • Recommendations • Questions • Considerations • More groups in WG1 • etc…..

  14. Workflowbetween WP 1 and WP-MRM -DeliverableOptions: Data Protein Gel band IVTT-system Protocols -Proteinvalidatedbythreelabs

  15. 25 proteinsfor MRM assays-USAL+UCM+Biogune→MRM WG • 25 novel proteins in both IVTT expressionsystems -USAL • 25 for DNA sequencingto UCM_ follow-up IVTT • Heavy proteins???? Planning-options-strategy • Quantity: amount 20 ug /average per protein HeLa IVTT as firstoption • 300 proteínas en 2 años--milestone

  16. Scouting • 500 Euros/proteinfor 200 ug • 200 Euros/protein full process (20 ugestimated) • 300 proteins total cost= 60000 Euros/2 years

  17. Antibodies

  18. http://dnasu.asu.edu/DNASU/GetVectorDetail.do?vectorid=157

More Related