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Hi André,

Patent is territorial. If the inventors of CN101864407A do not have a patent in your country, you are free to use the invention within your country. As academic researcher, you can also use an invention under "fair use". Each country may have different interpretations of "fair use". TEV has very good solubility, the mutations disclosed in CN101864407A are not needed to achieve high soluble expression.

In US, certain TEV proteases are protected by US5532142 and US7494786 patents. US5532142 is for a wild-type TEV protease. It has expired. Yale patent US7494786 covers some S219 mutations that most people use to stabilize TEV protease. NCI has an abandoned patent application for similar S219 mutations. Paying a fee to NCI does not avoid the infringement of US7494786. 

If you are concerned about patent infringement, TurboTEV is your choice. TurboTEV is Accealgen proprietary product that is not stabilized by any mutation. It has more robust activity than those widely used S219 mutants.

Hope this helps,

Chun

-----Original Message-----
From: André Mourão <[log in to unmask]> 
Sent: Friday, February 15, 2019 1:41 PM
To: [log in to unmask]
Cc: [log in to unmask]
Subject: Re: [ccp4bb] TEV Protease in low reducing agent?

Dear  Chun and all,

I do have a questions:
If I find the patent describing this mutation, (ttps://patents.google.com/patent/CN101864407A/en, maybe it is not), and If I reproduce those mutations in my lab, and If I express and purify these mutated protein and used it in my lab for academic research, does the lab that I represent have to pay anything, as this is a non profit mailing list?
looking forward to your answer.

Kind regards
André

----- Original Message -----
From: "Chun Luo" <[log in to unmask]>
To: [log in to unmask]
Sent: Wednesday, 13 February, 2019 21:37:27
Subject: Re: [ccp4bb] TEV Protease in low reducing agent?

Hi Nicola,

Our TurboTEV (http://accelagen.com/TurboTEV-datasheet.htm) is very stable and does not need additional reducing agent for its activity. The storage buffer has 1 mM TCEP. It'll be likely <0.1 mM in your digestion. You can get it from our European distributors or directly from Accealgen. Sigma probably sells it in Europe as well. TurboTEV is shipped with gel packs and can be stored at -20 for long time.

Cheers,
Chun
Accelagen

-----Original Message-----
From: CCP4 bulletin board <[log in to unmask]> On Behalf Of Nicola Evans
Sent: Wednesday, February 13, 2019 10:46 AM
To: [log in to unmask]
Subject: [ccp4bb] TEV Protease in low reducing agent?

I am interested in purifying proteins with 1 or 2 disulphide bonds, and have been using enterokinase to cleave off N-terminal tags but we have had issues with poor cleavage and want to try TEV cleavage instead. However TEV protease is usually kept in a high amount of DTT and I am concerned about reducing the disulphide bonds in my purified proteins. I have used TEV protease before with 0.25mM TCEP and it worked well. Is there a way to use TEV with very little or no reducing agent? Perhaps by optimising conditions such as adding glycerol? We were thinking of buying in commercial TEV protease so any advice on that is also welcomed (is there any merit, except cost, to making it ourselves?)

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