We found a small protein that we can aggregate, after purification. We do not believe them to be in their native state. So what properties of the elution/storage buffer can prevent/break protein (pure) aggregation. Does this have anything to do with the inherent properties of proteins? If yes, what are they? Can we understand the protein as such as we are not able to functionally characterize the protein at present, although we know the sequence characteristics. Where are we missing the catch, the monomer?
An attempt to answer some of your questions:
1. What properties of the elution/storage buffer can prevent aggregation? -->
Do you have enough reducing agent in your final purified protein vial?
2. Does this have anything to do with inherent properties of protein?If yes, what are they? --> It may be the case that your protein is highly hydrophobic?!! Try to predict the hydrophobic patches in your protein using sequence data that you have.
Hope it helps,
Best,
Sagar
Proteins containing Cys with free thiols may cross-link with each other over time if the concentration is high.
If cross-linking is not the problem, a 0.5 M solution of Arginine can help keep proteins folded
Another choice would be to add TMAO (trimethyl amine N-oxide) or glycerol.
A list of possible additives can be found in the review below. The exact additive you need will depend on its compatibility with the final assay.
Is the pI of the protein near neutral?
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