1 / 18

Microseeding Matrix Screening and other developments in automatic protein crystallization

Microseeding Matrix Screening and other developments in automatic protein crystallization Patrick Shaw Stewart Douglas Instruments Limited (near Oxford, UK): Peter Baldock, Patrick Shaw Stewart, James Smith. Microseeding in screening experiments. Allan D’Arcy Novartis, Basle

adamselizabeth
Download Presentation

Microseeding Matrix Screening and other developments in automatic protein crystallization

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. Microseeding Matrix Screening and other developments in automatic protein crystallization Patrick Shaw Stewart Douglas Instruments Limited (near Oxford, UK): Peter Baldock, Patrick Shaw Stewart, James Smith

  2. Microseeding in screening experiments Allan D’Arcy Novartis, Basle 2006 ‘Matrix-seeding script’ An automated microseed matrix-screening method for protein crystallization. ActaCryst. (2007). D63, 550–554

  3. Phase diagram of a protein precipitate nucleation [Protein] metastable zone clear [Precipitant]

  4. Microseeding in screening experiments Gregory Ireton and Barry Stoddard. 'Microseed matrix screening to improve crystals of yeast cytosine deaminase'. Acta Crystallographica section D60 (2004), 601–605. With Ca2+ conditions they couldn’t get crystals without microseeding.

  5. Phase diagram of a protein precipitate [Protein] metastable zone clear [Precipitant]

  6. Microseeding in screening experiments • May explain why there are SO many hits with MMS • Can be used to CONTROL number of conditions

  7. Microseeding in screening experiments James Smith 2006 ‘Matrix-seeding script’

  8. Microseeding in screening experiments James Smith 2006 ‘Matrix-seeding script’

  9. Microseedinginscreeningexperiments • Lesley Haire, NIMR: • 6 hits in a screen, all poorly formed. • Seeding - 30 hits, including several well-formed crystals. • Jens-Christian Navaro Poulsen, University of Copenhagen • 1 hit in 288 wells • Seeding - 10 hits in 96 wells • Using one of those crystals for a second-generation seeding experiment - another 10 hits in 96 wells

  10. Microseedinginscreeningexperiments Noseeds Laura Cendron University of Padova

  11. Microseedinginscreeningexperiments Noseeds Seeds Laura Cendron University of Padova

  12. Phase diagram of a protein [Protein] [Seeds]

  13. Mini-optimization

  14. Under £24,000 Under $46,000

  15. Bullet points: • 100 + 100 nl sitting drop • Only 9.8 μl of protein to set up a 96-well plate • Additive screens • Microseeding • Mini-optimization and optimization

  16. Jonathan Hadden • 96-well plate takes 8 minutes • There is an evaporation shield

  17. Under £24,000 Under $46,000

  18. ORYX (Arabian) The Biblical Zoo in Jerusalem

More Related