APPLICATION
Separation and purification experiment of recombinant protein GFP
2023-02-03
1. Affinity chromatography
Instrument recommendation |
SDA series |
SCG-P series |
Chromatography packing material: |
NW Rose Ni FF (M07D2203) CV:1mL |
|
Sample: |
GFP fermentation broth was crushed and centrifuged supernatant, loaded according to 20mg/mL load |
|
Buffer: |
Balance fluid: 20mM Tris-HCl 0.5M NaCl PH7.5 Eluent: 20mM Tris-HCl 0.5M NaCl PH7.5 Washing liquid 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 |
|
How to run: |
Equilibrium 5CV--Loading sample--Rinse 5CV--Elution 5CV--Elution①5CV—Elution②5CV--Equilibrium 5CV |
Recovery rate % |
Purity % |
|
Eluent |
90 |
83.4 |
2. Hydrophobic chromatography
Instrument recommendation |
SDA series |
SCG-P series![]() |
Chromatography packing material: |
UniHR Butyl 30L( L01W15KB) CV:1mL |
|
Sample: |
Affinity Eluent adjustment, Cond, pH are consistent with the balance solution, all samples are loaded |
|
Buffer: |
Balance fluid: 20mM Tris-HCl + (NH4)2SO4 PH7.5,Cond60ms/cm Eluent:20mM Tris-HCl Regeneration solution: 0.1M NaOH |
|
Flow rate: |
0.3ml/min |
|
How to run: |
Equilibrium 5CV--Loading sample--Rebalance 5CV--Elution 5CV--CIP5CV--Balance 5CV |
|
The detection yield of collected flow-through liquid was 74.7% |
Recovery rate % |
Purity % |
|
Eluent |
74.7 |
92.3 |
3. Desalination
Instrument recommendation |
SDA series |
SCG-P series![]() |
Chromatography packing material: |
G-25 CV:53mL |
|
Sample: |
GFP hydrophobic flow through solution |
|
Buffer: |
Balance fluid: 20mM Tris-HCl,PH7.5 |
|
Flow rate: |
0.3ml/min |
|
How to run: |
Equilibrium-loading-washing |
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