Root > Products > Strep-Tactin® & Strep-tag® > Strep-Tactin Resins > Superflow (high capacity) > Purification using Strep-Tactin Superflow and Superflow High Capacity

Comparison of Strep-Tactin® Superflow® vs. Superflow® high capacity

Purification of Strep-tag® II proteins via Strep-Tactin® Superflow:

Strep-Tactin® Superflow® for intact virus like particle (VLP) 4.8 MDa purification

Purification of intact virus like particle using Strep-Tactin Supeflow
Figure 1, left to right: M; Rainbow Marker RPN 755
GE Healthcare Biosciences Corp.,
1 and 2; different elution fractions of VLPs
fused to Strep-tag II after one-step Strep-Tactin
affinity purification from a crude lysate of E. coli.

1 liter culture was induced at an OD600 of 0.6 using anhydrotetracycline (AHT) and protein expression was performed at 37°C for 3 hours at 200rpm. Cells were then pelleted, resuspended in 20 ml Buffer W (100 mM Tris-Cl, 150 mM NaCl, 1mM EDTA) and sonicated. The insoluble material was pelleted and the crude lysate was loaded on a Strep-Tactin® Superflow® column at a flow rate of 1ml/minute. After washing off the host proteins, the VLPs fused to Strep-tag® II were eluted using 2.5mM desthiobiotin in Buffer W. pASK-IBA7 was used for cloning.

For the purification of intact VLPs, the careful choice of both an ideal affinity tag system and the resin support is important:

  • The affinity chromatography process should be gentle in order to keep the VLPs intact, as with Strep-tag®.
  • The resin support packing within the column must allow penetration of the large VLPs. 

While MacroPrep® carriers did not meet criterion (2), since the viral particles accumulated on top of the column, Sepharose® and Superflow® enabled purification. Superflow® can be used in FPLC systems, whereas Sepharose® is only suitable for gravity flow purification.

These results were kindly provided by L. Stöckl and B. Brandenburg, Robert-Koch-Institute, Berlin.
Brandenburg B., Stockl L., Gutzeit C., Roos M., Lupberger J., Schwartlander R., Gelderblom H., Sauer I. M., Hofschneider P. H. and Hildt E. (2005)
A Novel System for Efficient Gene Transfer Into Primary Human Hepatocytes Via Cell-Permeable Hepatitis B Virus–like Particle, HEPATOLOGY (42),1300-1309