How to Separate recombinant Enterokinase and Aim Protein？
March 8, 2022
Designed Recombinant Enterokinase can specially cut recombinant fusion protein, including two types: one is REK-B, it is label free, a native sequence of bovine enterokinase light chain; another is REK-H, contain 6×His tag.
Yaxinbio can specially supply label - free REK-B, with a higher specific activity around 10000 Unit/mg pro. According to the definition of the activity, 1 Unit recombinant enterokinase can cut 0.5mg aimed protein, the content of recombinant enterokinase in digestion solution is ten thousandth (0.01%). Therefore, it is very low, and not necessary to consider the impact of REK on downstream results. It is not recommended to separate REK from the digestion solution specially.
If you do need to remove REK, recommend removal method is as follows:
1) Trypsin inhibitor affinity gel is recommended. As recombinant enterokinase is a serine protease, trypsin inhibitor can be combined with enterokinase so as to achieve the purification purpose.
2) It is also proposed to use ion-exchange chromatography to remove small amount of recombinant enterokinase.
At pH 8.0, recombinant enterokinase can combine with DEAE gel,and then It can be eluted by 0.15M NaCl.