Semi-preparative purification within minutes
Pre-packed column with Strep-Tactin POROS and accessories.
The green fluorescent protein (GFP) with C-terminal Strep-tag II was expressed in the cytoplasm of E. coli. After sterile filtration the crude lysate was separated via Perfusion Chromatography® on Strep-Tactin POROS 20 (5 mg/ml) under physiological conditions (100 mM Tris-Cl pH 8.0, 1 mM EDTA). Elution was performed by the addition of 5 mM desthiobiotin in the same buffer and chromatography was monitored by measuring the absorbance of the eluate at 280 nm.
The purification is documented on a coomassie stained SDS gel where samples from the crude lysate (lane 1) and from the elution peak at 6.7 minutes (lane 2) had been applied. 3 mg GFP could be obtained in one step when running a 1.7 ml column on a BioCAD® work-station.
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