While I understand that you want to have protein concentration at its 
solubility limit, i had several proteins which can go to 60-80 mg/ml, they 
seems to get crystallized at 10-15 mg/ml. All you want is saturation  in 
crystallization conditions.
Dhiraj srivastava

________________________________
From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of Jon Cooper 
<0000488a26d62010-dmarc-requ...@jiscmail.ac.uk>
Sent: Monday, February 5, 2024 5:32 AM
To: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK>
Subject: [External] Re: [ccp4bb] Crystallizing a tough target

Limited proteolytic might help, too.

https://www.nature.com/articles/nmeth1118


Best wishes, Jon Cooper. jon.b.coo...@protonmail.com

Sent from Proton Mail mobile



-------- Original Message --------
On 5 Feb 2024, 10:27, kavyashreem < kavyashr...@instem.res.in> wrote:


Dear All,

Has anyone worked on a protein which is highly soluble even at 80mg/ml?

We have one such candidate, which does not precipitate even at 80mg/ml instead 
forms phase separated globules in crystallization plate, which eventually 
hardens over a period of 1 to 1.5 months (which is florescent under UV 
microscope.)

We tried screening at different pH, but failed to get any hits.

Since we got few conditions in which the phase separated globules solidified, 
we focused on them and expanded with 120mg/ml protein, still there were not 
visible precipitates except for the phase separation. This has been a 
challenging target so far. We have tried with different constructs, which 
unfortunately are not soluble!

Does POMs help in such cases? Or do you have any other suggestion.

Thank you

Regards

Kavya




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