蛋白质层析技术

1、HV (flow rate) 1) Thyroglobulin 2) Gamma-globulin 3) Beta-globulin 4) Cytochrome C50-100 m UNOsphere30-60 mMacro-Prep 5020-40 mMacro-prep 2510 mBio-ScaleUNO 5-10 m beads 1-20 ml columns High operating pressures, (80 psi) 20-30 m beads 20-500 ml Medium operating pressures (30-100 psi) 40-200 m beads

2、20 ml-1000 liters Low to medium operating pressures (5-50 psi)IEXHICHydroxyapatiteAffinitySECAmmoniumsulfateHICHydroxyapatiteAffinitySECHICHydroxyapatiteIEXAffinitySEC Product Properties Stability Charge at different pH Hydrophobicity Molecular Size Essential co-factors Feed PropertiesCritical impur

3、itiesProduct concentrationIntroduced contaminants Final Product specifications Purity Cost per unit Lot sizeSeparation ChemistryMethod ScoutingChemical Stability (lifetime, hygiene) ProductivityPurity, Final Scale, EconomicsChromatography media with required process performanceBase MatrixBead SizeMo

4、del 2110RediFracFrac 200BioFracPerformancePerformanceGilson 204PricePriceFrac 100$1000.00 US List List PricePerformance01002003004005006000.0000.0050.0100.0150.0200.025Volume(ml)OD280nm UV050100150200250HIC for FABPConductivity(mS/cm) ConductivityNVW5541 22./HLN020406080100120-0.020.000.020.040.060.

5、080.100.120.14Volume(ml)OD280nm(AUFS) UV20406080100hpag4 on Superdex200 Con.(mS/cm) Conductivity-20246810 12 14 16 18 20 22 24 26 28 30 32 340.00.10.20.30.4hpag4 on resourceQVolume(ml)OD280nm(AUFS) UV020406080100Con.(mS/cm) Conductivity-500501001502002503003504000.00.51.01.52.0TIMP on MacroPrepHighS

6、Volume(ml)OD280nm(AUFS) UV020406080Con.(mS/cm) Conductivity0204060801000.00.10.20.30.40.50.6TIMP and HW on Superdex200Volume(ml)OD280nm(AUFS) UV20406080100120140Con.(mS/cm) Conductivity-2024681012141618202224260.000.020.040.060.080.10TIMP on MonoQVolume(ml)OD280nm(AUFS) UV01020304050Con.(mS/cm) Cond

7、uctivity0102030400.000.010.020.030.040.050.06HM on souce15ISOVolume(ml)OD280nm(AUFS) UV050100150200250Con.(mS/cm) Conductivity-200204060801001201401600.00.10.20.30.40.50.6Volume(ml)OD280nm UV050100150200Size-exclusion for FABPConductivity(mS/cm) Conductivity01002003004005006000.0000.0050.0100.0150.0

8、200.025Volume(ml)OD280nm UV050100150200250HIC for FABPConductivity(mS/cm) Conductivity-1001020304050607080-0.10.00.10.20.30.4Ion-exchange for FABPVolume(ml)OD280nm UV020406080100Con.(mS/cm) Conductivity010203040500.00.10.20.3Volume(ml)OD280nm(AUFS) UV020406080100GST on BioscaleQ2Con.(mS/cm) Conductivity020406080100120-0.10.00.10.20.30.4GST with 2mMDTT on Superdex200Volume(ml)OD280nm(AUFS) UV20406080100Con.(mS