We have just created a therapeutic protein in the laboratory

We have just created a therapeutic protein in the laboratory that can cure Malaria. We have successfully expressed the protein in E. coli in high quantities and need to purify it for further analysis. The primary sequence of this therapeutic protein is shown below using the 1 letter abbreviation for each amino acid. mmnmkivlfs lllfvlrwni iscnkndknq gvdmnvlnny enlfkfvkce ycnehhhhhh hhhhhhhhhh Based on the primary sequence given above what type of chromatography should be used to purify this protein? Describe the steps of how you would use the chromatography method that you chose in part A to purify this protein (this answer should convince me that you understand how the chromatography works, the outcome, etc ... Draw pictures if needed).

Solution

a) your protien seems to contain a poly histidine or a his-tag hence affinity chromatography with nickel-cobalt resin would be most appropriate choice to purify the protien ...

b) we will add bacterial lysate through chromatographic column the protien of intrest would bind to Nickel OR Cobalt then after phosphate wash through resin most of non specific protiens will wash away ... finally in the last step we will add imadazole which will compete with histidine imadizole and elute the protien fraction of intrest

Hope it helps...... Feel free to discuss more with me ... I will be glad to help

Regards