Purifications of HTT and HTT-HAP40 from insect (Sf9) and mammalian (EXPI293F) expression systems

I am really excited to be working in collaboration with so many great groups in the Huntington’s disease research space. One of my contributions to many of these collaborations is to provide huntingtin (HTT) and HTT-HAP40 protein samples with different polyglutamine tract lengths. My collaborators can then use the material to investigate both the structure and function of the HTT protein and see is there are any different characteristics for the unexpanded and expanded polyglutamine HTT protein species.

I primarily am using protein made in insect cells or Sf9. This is our favoured system as it is scalable and relatively inexpensive so we can rapidly make lots of protein sample. However, we also make HTT samples in a human-derived cell line called EXPI293F. This should (!) produce a protein sample which is more representative of the protein we find in our own cells. Comparing the characteristics of HTT made in different systems is an ongoing project.

Recently I have made a number of different samples in Sf9 and EXPI293F cells, both HTT and HTT-HAP40. These are to be shipped off to labs around the world for different experiments which I think is very exciting!

Gel filtration and SDS-PAGE analysis of HTT-HAP40 Q23 showing a highly pure final sample.

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