I am working on Differential Scanning Calorimetry (DSC) for my protein. Mainly I want to check its melting temperature and enthalpy changes. I first tried two standard proteins ( BSA and lysozyme) and it works fine. However, I found aggregation after experiments for my protein. The concentration of my protein is 1.5mg/ml and the buffer is 10mM Phosphate buffer with 100mM NaCl at pH7.0. Is there any way to avoid aggregation during DSC? Thanks!
Different proteins behave differently under the same experimental conditions.
You must test different conditions (pHs, ionic strengths, buffers, additives...) and evaluate aggregation. You may use DSC as a diagnostic tool, or employ other biophysical tools (e.g. DLS after temperature treatment or with temperature scanning).
On the other hand, protein aggregation may be not relevant for protein stability evaluation if the aggregation occurs at much higher temperature compared to the protein unfolding process (that is, if protein unfolding and protein aggregation are rather uncoupled processes). Unfolded protein at high temperature is prone to aggregation.
Of course, it is because of the denaturation of protein. In fact, you are measuring the denaturation and protein aggregates after the denaturation.
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