protein endotoxin free- kit and system advice

HI....

we have one project where we should remove LPS from purified proteins that we produce in e.coli for further anaylsis in human cells.

we are using the ToxinEraserTM Endotoxin Removal Kit form GeneScript and Pierce® High-Capacity Endotoxin Removal Resin from Thermo.

Which kit are you using?

I have a friend who points my attention to CleanColi Expression System.

It is for protein expression and contains a genetically modified lipopolysaccharide that does not create an endotoxic response in human cells. Because of this, researchers will be able to increase their productivity since they will no longer need to remove unwanted endoxin, Lucigen and RCT said.

do anybody have experiments with this system?

thanks in advance,

Carlo

 

Posted on 03-May-2013 11:22 CEST
Carlo Carolis

Dear Carlo,

The CleanColi Expression System seems interesting. 


If you decide to stay with E. coli you might want to have a look at these publications to reduce endotoxin levels during the purification of His-tagged and GST-tagged proteins produced in E coli.

Single step protocol to purify recombinant proteins with low endotoxin contents. Reichelt P, Schwarz and Donzeau M. Protein Expression and Purification 46, 483-488 (2006).


Simultaneous metal chelate affinity purification and endotoxin clearance of recombinant antibody fragments. Zimmerman T, Petit Frère C, Satzger M, Raba M, Weisbach M, Döhn K, Popp A and Donzeau M. Journal of Immunological Methods 314, 67-73 (2006)

Best wishes.

Patrick

Posted on 03-May-2013 14:39 CEST
Patrick Duriez

Hi:

I have very good experience with  HYGLOS  EndoTrap® blue and EndoTrap® red

You can see some information on: http://wolfson.huji.ac.il/purification/PDF/Endotoxin/PROFOS_EndoTrapBlue.pdf

 

Mario

Posted on 05-May-2013 17:11 CEST
Mario Lebendiker

thanks a lot for the information!!!

Posted on 23-May-2013 11:59 CEST
Carlo Carolis