PUREfrex® ver. 1.0 vs 2.0

Overview

PUREfrex® was launched in 2011, now we improved the protein productivity of PUREfrex® and named the advanced version “PUREfrex®2.0”.

We modified the preparation methods of all components that were purified from E.coli and optimized the factors’ composition. As a result of this modification, PUREfrex®2.0 achieved 2-10 times higher protein productivity than the previous version.

Furthermore, the yield of active form of protein which needs a proper folding to active was increased when synthesized in PUREfrex®2.0 with chaperon and/or disulfide bond isomerase.

SDS-PAGE

PUREfrex® 1.0 PUREfrex® 2.0
The amount of DHFR production per one kit (250 μL) 40 μg 150 μg
How many kits need for 100μg of DHFR production? 660 μL (> 3kits) 160 μL (< 1kit)
The amount of GFP production per one kit (250 μL) 25 μg 200 μg
How many kits need for 100μg of GFP production? 1000 μL (> 4 kits) 125 μL (< 1kit)

Compare the activity of synthesized protein

GFP

Fluorescence of synthesized with PUREfrex® 2.0 is more than 10 times higher than PUREfrex® 1.0.

GFP activitySynthesized GFP

 

Acid phosphatase (AppA)

AppA from E.coli with 5 disulfide bonds, 1 of which is non-consecutive. The enzyme activity of synthesized AppA is increased when AppA synthesized with PUREfrex® 2.0 and DS supplement which contains disulfide bond isomerase DsbC.

AppA activity

 

Luciferase

The enzyme activity if luciferase is increased when luciferase synthesized with PUREfrex 2.0 and Dnak mix which contains DnaK, DnaJ and GrpE.

Luciferase activity

 

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