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The US patent for Protease Pro 39

On 13 March 2007, The United States Patent and Trademark Office issued patent No US7189540 

“OPTIMIZED RECOGNITION SITE OF THE ALPHAVIRUS NON-STRUCTURAL PROTEASE FOR TAG REMOVAL AND SPECIFIC PROCESSING OF RECOMBINANT PROTEINS”  to Icosagen AS (formerly Quattromed AS).

Pro39 cleaves amino acid sequence DVLRLGRAGA?G at the N-terminus or DVLRLGRAGA?XIFSS at the C-terminus, where X is preferably Y or G but, can be any amino acid except P.
The unit characterising the activity of the enzyme is defined as follows: 1 unit of Pro39 cuts 10 µg of test proteins with an effectiveness of 90% in a period of 1 hour at a temperature of 30 ºC.

The potential areas of use for Pro39 are:
• Removal of a short peptide sequence or tag added to recombinant proteins.
• Cutting of multi-domain proteins between domains.
• Processing of recombinant proteins under in vivo conditions.

During the cleaning of recombinant proteins either a short peptide sequence or tag, or some other easily cleaned protein, is frequently added to the protein being produced. The obtained proteins are cleaned via the tag or the easily cleaned protein with the aid of affinity chromatography. Since this “additive” containing protein is not suitable for all implementations, the tag or added protein must be removed. In order to do so a suitable protease recognition area is injected into the proteins or between the protein and tag, and after the cleaning of the “additive” containing protein the protein is processed with protease.
Pro39 is Semliki Forest virus (SFV) non structure protein nsP2 protease domain (341 aa, 3068 – 5541), which, in its nature, is a virus specific cysteine protease. Pro39 effectively cuts one of the three cutting sites located on SFV nonstructural polyprotein. Therefore, protease Pro39 depicts itself as an active and specific enzyme, which can be used in biotechnology and molecular biology research.
Research for the given patent application was done as joint work between scientists at Icosagen (formely Quattromed) and the Institute of Technology of the University of Tartu.

Pro39 is in the Icosagen product list as two different articles:
500 units (catalogue number Y1-100-005)
2500 units (catalogue number Y1-100-025)