Microfluidic Dialysis Protein Crystallization Jiang Huang GN Biosystems, Inc. March 26, 2009
Dialysis Protein Crystallization Method Pro: scans a wide concentration range, the reagent composition can be easily altered during the course of the experiment. Con: difficult to setup, protein consumption too high (5 to 350l/rxn), not HT compatible.
MICROBATCH HANGING DROP FD Venn Diagrams 3 1 Method Hits VDX 20 MB 8 FD MICROBATCH HANGING DROP Method Hits VDX 9 MB 15 FD 7 FD MICROBATCH HANGING DROP Method Hits VDX 1 MB 10 FD 5 FD Lysozyme Glucose isomerase Catalase
Microfluidic Dialysis Plate - Design open bottom microtiter plate protein inlet film dialysis membrane discs microfluidic plate adhesive sealing tape
protein inlet vacuum port dialysis membrane reagent well Microfluidic Dialysis Plate - Design adhesive film
Microfluidic Dialysis Plate Design Top View Dialysis chambers dimensions: Screening plate:18nl per chamber (240m dia., 400m deep). Optimization plate: 80nl per chamber (500m dia., 400m deep). Growth plate: 1l per chamber (1.6mm dia., 400m deep) Bottom View Vacuum port Dialysis chamber Microfluidic channel Protein port
Microfluidic Dialysis Plate – Sample Loading
FD Method Highlights Protein consumption as low as 15nl per dialysis chamber Fast set-up in as little as 4 minutes for 96 or 384 dialysis chambers Low capital equipment costs to begin running experiments High-throughput compatibility with standard dispensing robotics Easy translation and scale-up designs Chemically compatible with commercial reagent kits The dialysis membrane can be conveniently to allow easy loop access for crystal manipulation
FD Method Highlights Protein consumption as low as 15nl per dialysis chamber Fast set-up in as little as 4 minutes for 96 or 384 dialysis chambers Low capital equipment costs to begin running experiments High-throughput compatibility with standard dispensing robotics Easy translation and scale-up designs Chemically compatible with commercial reagent kits The dialysis membrane can be conveniently to allow easy loop access for crystal manipulation
FD Method Highlights Protein consumption as low as 15nl per dialysis chamber Fast set-up in as little as 4 minutes for 96 or 384 dialysis chambers Low capital equipment costs to begin running experiments High-throughput compatibility with standard dispensing robotics Easy translation and scale-up designs Chemically compatible with commercial reagent kits The dialysis membrane can be conveniently to allow easy loop access for crystal manipulation
Equipment and Accessories Needed 1.Vacuum pump or house vacuum with an ultimate vacuum ≤ 0.1mmHg *Air bubbles in every dialysis chamber will result due to insufficient vacuum A List of Qualified Vacuum Pumps at under $2,000 ManufactureModel#Ultimate Vacuum BOC/EdwardsRV31×10 -6 mmHg BOC/EdwardsEVA ×10 -3 mmHg Welch1400B-011×10 -4 mmHg Welch1399B mmHg BrinkmannV mmHg Thermo-Electron ×10 -3 mmHg Thermo-Electron ×10 -4 mmHg
FD Method Highlights Protein consumption as low as 15nl per dialysis chamber Fast set-up in as little as 4 minutes for 96 or 384 dialysis chambers Low capital equipment costs to begin running experiments High-throughput compatibility with standard dispensing robotics Easy translation and scale-up designs Chemically compatible with commercial reagent kits The dialysis membrane can be conveniently to allow easy loop access for crystal manipulation
FD Method Highlights Protein consumption as low as 15nl per dialysis chamber Fast set-up in as little as 4 minutes for 96 or 384 dialysis chambers Low capital equipment costs to begin running experiments High-throughput compatibility with standard dispensing robotics Easy translation and scale-up designs Chemically compatible with commercial reagent kits The dialysis membrane can be conveniently to allow easy loop access for crystal manipulation
via diameter: 1.2mm (570nl volume)via diameter: 0.3mm (20nl volume) crystal size: up to 500m longcrystal size: up to 50m long 50mg/ml Lysozyme vs. HCS1 #10 Translation and Scale-up screeningoptimizationgrowth dia.=240mdia.=500m dia.=1.6mm depth=400mdepth=400mdepth=400m
#2 #6 #7 #9 #13 #14 15 l dialysis bottom Optimization Plate Screening Plate Hampton Crystal Screen I 90x 20x Translation and Scale-up
FD Method Highlights Protein consumption as low as 15nl per dialysis chamber Fast set-up in as little as 4 minutes for 96 or 384 dialysis chambers Low capital equipment costs to begin running experiments High-throughput compatibility with standard dispensing robotics Easy translation and scale-up designs Chemically compatible with commercial reagent kits The dialysis membrane can be conveniently to allow easy loop access for crystal manipulation
FD Method Highlights Chemically compatible with commercial reagent kits (materials used: PMMA, epoxy, dialysis membrane) M. W. Toepke, D. J. Beebe, PDMS absorption of small molecules and consequences in microfluidic applications, Lab Chip, 2006, 6:
FD Method Highlights Protein consumption as low as 15nl per dialysis chamber Fast set-up in as little as 4 minutes for 96 or 384 dialysis chambers Low capital equipment costs to begin running experiments High-throughput compatibility with standard dispensing robotics Easy translation and scale-up designs Chemically compatible with commercial reagent kits The dialysis membrane can be conveniently to allow easy loop access for crystal manipulation
protein inlet vacuum port dialysis membrane reagent well Microfluidic Dialysis Plate - Design adhesive film