Optimisation of huntingtin domain protein sample buffer conditions

Previously, I wrote about how I successfully made a small chunk of the huntingtin protein. However, I then found that the sample was not very thermally stable. Therefore I have tried to improve the protein stability by looking at different solution conditions for the protein molecules by screening different buffers, pH, salt concentrations as well as the addition of additives to the sample.

The protein has a very high expected pI (this is the pH at which the protein has neutral overall charge) of pH ~ 9. Therefore it is not unexpected that the data show that moving away from this pH into more acidic conditions, stabilises the sample. Additives have little effect on the protein sample but glycerol is stabilising. All of the results from the buffer screening can be found on Zenodo.

Shown are the calculated Tagg (the temperature at which the protein aggregates in solution in °C) for each condition

I hope to next purify the protein in more acidic conditions starting tomorrow! I hope this may increase the yield and quality of the sample.

2 thoughts on “Optimisation of huntingtin domain protein sample buffer conditions

  1. I am not a scientist, I just read a lot. I just wanted to say, that I absolutely love you! Thank you for all you do Dr. Harding!

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