It really depends from the nature of the protein and if is
oligomerizing/agregating/forming polymers if any of this is reversible.
On the other side if you are working with one of the fibril forming
protein it will require optimizaiton on its on as they will form
naturally polymers.
Do you observed different specises when you analyze your protein by SEC
or if you are able to perfom DLS?
Additional information regarding your protein will be really helpful for
more detalied suggestions how to overcome your protein.
Best,
Nikolay
Nikolay Dobrev
Scientific Officer, Protein Expression and Purification Core Facility
EMBL Heidelberg, Meyerhofstraße 1, 69117 Heidelberg, Germany
T +49 6221 387 8633 | M +49 173 684 0532
twitter.com/EMBLorg | facebook.com/embl.org | youtube.com/user/emblmedia
Visit www.embl.org/events for a complete list of all EMBL events.
On 19/07/2020 14:15, S. Mohanty wrote:
Keep the protein concentration low during purification steps along
with using other anti-aggregation agent/s. Make sure that the pH at
which you are purifying is not close to the pI of the protein. Until
completely purified, all purification steps should be performed in a
cold room if it is a soluble protein.
Smita
On Sunday, July 19, 2020, 04:28:45 AM CDT, Sorin Draga
<sor.dr...@gmail.com> wrote:
I am not sure what you mean by polymer formation. Presuming that you
have optimized your protein concentration, pH and salt concentration,
you could try arginine as an anti-aggregation agent in your
purification (I presume you do FPLC). Have a look at chaotropic agents
used in protein purification, The answer is generally dependent on the
protein/proteins you are trying to purify and is not
necessarily straightforward.
Kinds regards
On Sun, Jul 19, 2020 at 12:08 PM 张士军 <21620150150...@stu.xmu.edu.cn
<mailto:21620150150...@stu.xmu.edu.cn>> wrote:
Dear all:
Any ideas to decrease protein polymer formation? my protein was
easy to form oligomers and precipitation when do purification,I
have tried add glycerol and DTT,both didn't work. Does anyone has
experience to avoid it happening. Thanks!
Best Regards
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Nikolay Dobrev
Scientific Officer, Protein Expression and Purification Core Facility
EMBL Heidelberg, Meyerhofstraße 1, 69117 Heidelberg, Germany
T +49 6221 387 8633 | M +49 173 684 0532
twitter.com/EMBLorg | facebook.com/embl.org | youtube.com/user/emblmedia
Visit www.embl.org/events for a complete list of all EMBL events.
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