1. Affinity chromatography
Recommended instrument | SDA /SCG -P Series |
Chromatographic packing | NW Rose Ni FF (M07D2203) CV:1mL |
Sample | GFP fermentation liquid was crushed and centrifuged supernatant, and the sample was taken according to 20mg/mL load |
Buffer | Equilibrium liquid: 20mM Tris-HCl 0.5M NaCl pH7.5 Eluent: 20mM Tris-HCl 0.5M NaCl pH7.5 Detergent 1:20mM Tris-HCl 0.5M NaCl 0.5M imidazole pH7.5，20% Eluent 1: 20mM Tris-HCl 0.5M NaCl 0.5M imidazole pH7.5，55% Eluent 2: 20mM Tris-HCl 0.5M NaCl 0.5M imidazole pH7.5 |
Flow rate | 0.3mL/min |
Operation method | Balanced 5CV-- Loading sample -- Eluting 5CV-- Cleaning 5CV-- Elution ①5CV -- Elution ②5CV-- Balanced 5CV |
| Recovery rate% | Purity% |
Eluent | 90 | 83.4 |
2. Hydrophobic chromatography
Recommended instrument | SDA /SCG -P Series |
Chromatographic packing | UniHR Butyl 30L( L01W15KB) CV:1mL |
Sample | The concentration of the eluent was adjusted, Cond and pH values were consistent with the equilibrium solution, and all samples were uploaded. |
Buffer | Equilibrium liquid: 20mM Tris-HCl + (NH4)2SO4 pH7.5, Cond 60ms/cm Eluent : 20mM Tris-HCl Regenerated liquid: 0.1M NaOH |
Flow rate | 0.3ml/min |
Operation method | Balance 5CV-- Loading sample -- Rebalance 5CV-- Elution 5CV--CIP 5CV-- Balance 5CV |
74.7% yield of collected flow-through liquid assay |
| Recovery rate% | Purity% |
Flowthrough | 74.7 | 92.3 |
3. Desalinize
Recommended instrument | SDA /SCG -P Series |
Chromatographic packing | UniHR Butyl 30L( L01W15KB) CV:1mL |
Sample | The concentration of the eluent was adjusted, Cond and pH values were consistent with the equilibrium solution, and all samples were uploaded. |
Buffer | Equilibrium liquid: 20mM Tris-HCl + (NH4)2SO4 pH7.5, Cond 60ms/cm Eluent : 20mM Tris-HCl Regenerated liquid: 0.1M NaOH |
Flow rate | 0.3ml/min |
Operation method | Balance 5CV-- Loading sample -- Rebalance 5CV-- Elution 5CV--CIP 5CV-- Balance 5CV |
74.7% yield of collected flow-through liquid assay |
