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Trypsin Expression and Purification


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#1 element771

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Posted 29 July 2009 - 11:21 AM

Is there any way to express and purify recombinant trypsin or chymotrypsin?

I am also worried about autolysis.

In other words....

Can anyone point to references or protocols that would allow for expression and purification of trypsin while taking measures to prevent autolysis?

Thanks a ton!

#2 klinmed

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Posted 29 July 2009 - 01:28 PM

View Postelement771, on Jul 29 2009, 09:21 PM, said:

Is there any way to express and purify recombinant trypsin or chymotrypsin?

I am also worried about autolysis.

In other words....

Can anyone point to references or protocols that would allow for expression and purification of trypsin while taking measures to prevent autolysis?

Thanks a ton!


You should look at US patent 7351549 "Method for the manufacture of recombinant trypsin".

Check out the US Patent Office web site, you can find the complete application there.

Hope this helps

#3 T C

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Posted 30 July 2009 - 06:55 AM

Hey,

I generally resuspend trypsin in water so that there is no autolysis and in the digests, add buffer.

Best,
TC

View Postelement771, on Jul 30 2009, 01:51 AM, said:

Can anyone point to references or protocols that would allow for expression and purification of trypsin while taking measures to prevent autolysis?


#4 element771

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Posted 30 July 2009 - 07:15 AM

View PostT C, on Jul 30 2009, 07:55 AM, said:

Hey,

I generally resuspend trypsin in water so that there is no autolysis and in the digests, add buffer.

Best,
TC

View Postelement771, on Jul 30 2009, 01:51 AM, said:

Can anyone point to references or protocols that would allow for expression and purification of trypsin while taking measures to prevent autolysis?



The problem with this is that I need to isotopically label the trysin. Thanks though.





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