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Microplates for High Throughput Crystallization Jeri McMahon Corning Life Sciences Europe
Central Dogma in Life Sciences DNA RNA Protein Structure/Function HTC plates HIGH THROUGHPUT DISCOVERY PLATFORM Target Id Assay Develop. Screening Lead Optimization Strategic Importance to Corning Structural Genomics/Proteomics Structure based Drug Discovery
Ready for Prime Time? HTC Robots Robotic trays ?
HT Crystallization Market HTC SBDD Structural Genomics
Segmentation: HT Crystallization Market • Structure Genomics Centers 7, 150 millionx2 • Biotech & Pharma: Core Labs • Individual Crystallography Labs: 2000 WW • Structure Genomics Company: Syrrx, SGX, Astex, PSF
Crystallization methods: Hanging Drop Sitting Drop CCP Cat. #3773 Costar 3424 plate
Crystallization methods: Sandwich Drop MicroBatch Membrane Protein? 96 Well Costar Plate™ 96 well V Btm, Clear, PS 3367 single, RB, PS, S 3795 100/Cs, RB, PS, S
HTC plates: 96 Well Sitting Drop: Corning Greiner A. Robbins 384 Well Sitting Drop Corning 96 Well Hanging drop: Neuroprobe Corning, --coming in 2004
1st 96 well sitting drop: 1997/1998? The first 96 Well Sitting Drop Pre-SBS, not low robotic compatibility
HT sitting drop: 2001 Greiner 96 1:3 Square reservoir 3 square protein well with flat btm, PS
HT sitting drop: 2001 Corning 96 1:1 Novel merged well, round reservoir, round protein well with R btm *Patent Pending
96 well Corning Sitting Drop Plate: • Simple 1:1 configuration sitting drop: • One large reservoir well and a small protein well Protein Well 10 uL Reservoir Well 210 uL
Manual: with multi-channel pipettors • Robotic: SBS standards for automation, • require 96 not 1536 well precision robots • Efficient vapor space: • with Novel merged well design • Round well with Conical bottom: evolved from 24well sitting drop plate • Polarization free protein well Product Features:
5 “First” brought into the field by CCP? First 96 well sitting drop plate with round well First 96well sitting drop plate with conical bottom First sitting-drop plate with low polarization First sitting-drop plate with Ultra-low liquid loss First optical quality sitting-drop plate high chemical resistance: Acetone, acetic acid, butanone, ethanol, isopropanol, methanol, DMSO, nitric acid (65%), sulphuric acid (40%), hydrogen chloride (36%), and 33% ammonia. Corning Plate other plate Plate wells as viewed under polarized light
Fast Drops Manual: 20 hrs < 3 days 10 Proteins x 800 Conditions in half a day Traditional 130 hrs >3 weeks 1 2 3 4 5 6 7 8 9 10 Ten Distinct Proteins or Protein- Ligand Complexes Hampton Screens I & II 100 Wizards I & II 100 Greed Screens 200 Random 400 (Crystool) 400 Total Reagents in Crystal Screen 800 Automated Fast Drops: 3 hrs! 10 X 800 = 8,000 drops
HT sitting drop: 2002 Inteli PlateTM9 6 by Art Robins Enterprises.
HT sitting drop: CCP 384 2002 Corning CCP 384 1, 1st HTC plate to have all SBS well locations: 384 2, Has the highest # of HTC screening conditions: 192
Simple 1:1 Configuration Sitting Drop Protein Well 3.4 uL superb low polarization viewing window Reservoir 105 ul No, not a bottom less plate :-)
CCP 384 Top features: • The only crystallization plate built to meet full SBS standards including well locations • Optimized for robotic HT sitting-drop protein crystallization • Features large square reservoirs with round protein wells • Reservoir well is 105 µL, ideal for loading 25-75 µ l screens • Protein well volume is 3.4 µL, ideal for 1.5+1.5, 1+1, 0.5+0.5… • Made of the same advanced optical polymer as the original CCP
CCP 384 Test Drive with Tecan Genesis
HT sitting drop: 2003, Coming soon CCP II Conical flat bottom, Designed for 1-2 ul drops. Sloped for easy Crystal Access Smaller protein well size
Trends: Chemical resistance will become more important as screen conditions grow CCP & CCP 384 Compatibility Table: Acetic Acid 100% Nitric Acid 65% Sulphuic acid 40% HCL 36% Ammonia Solution 33% DMSO Acetone Butanone Methanol Ethanol Isopropanol
Trends (Unmet needs in the HTC market) • Coated plates for anti static • Differential treated baby well for auto-centering of protein drops • HTC plates for direct X-ray • SBDS plates • Open Platform Hanging Drop 96 • Microfluidic Hanging Drop Plate • Trans Drop TM Plates • Hanging Drop 384 • Crystallization Kits??? & so on & so on….
Universal Optical Sealing Tape • The tape is pressure-sensitive. Customers can use an applicator or roller to apply the tape. • Potential uses in other applications • Key features of the sealing tape: • High optical quality for microscope observation of protein crystals on plate • Minimal evaporation of protein well content • Minimal cross-well contamination • Suitable for use between –70 and 100°C • Compatible with aqueous solutions and organic solvents
Where to get them? Fisher, VWR, Hampton R.
HTC plates in the future: • Corning 96 Well HT Hanging Drop Plates • Grease free • Polarization free • Easy Crystal Access • Compatible with Cyberlab 240 Hanging Drop Robot • Compatible with All sitting drop robots • Priced similar to Sitting Drop Plates • ---Coming in 2004, Patent Pending