Separation and purification experiment of recombinant protein GFP

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

Instrument recommendation

SDA 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

Instrument recommendation

SDA 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