Hi, dr

If your desired protein size less then 5kda then you can try TFF (10kda) in flowthrough (permeate mode), Endotoxin have higher size so endotoxin will retain in retenate and your product will come in flow-through.

Hope this is useful.

Hi Paul,

What is the model you are using? If using tissue culture you can manipulate both the host and/or pathogen to remove LPS driven effects. For example using TLR4 KO or caspase-11/4 KO will reduce LPS driven signalling and pyroptosis. Alternatively, depending on the pathogen, mutants are available in key steps of lipid A biosynthesis eg MsbB in Salmonella. Each would have to be carefully controlled for and checked in the literature as these mutants tend to have growth defects.

Good luck

Mitch

Hi,

You could try using commercial columns such as EndoTrap. Acoording to their Internet site, EndoTrap is an affinity chromatography resin based on a bacteriophage-derived protein which binds to endotoxins with high affinity and specificity. The EndoTrap protein captures the conserved region of the inner core of LPS molecules and is thereby able to bind to all kind of endotoxins from Gram-negative bacteria. It is neither an antibody, a synthetic peptide nor Polymyxin B-based.

Best wishes,

Jean-Paul

Hi Paul,

I agree with the above recommendation of EndoTrap columns (or bulk resin) to remove the LPS. There is no contamination of your sample and, in fact, this can be confirmed with the EndoTrap Leakage ELISA after running it though the column. There are two types of EndoTrap columns, but the EndoTrap HD is the most efficient at removing endotoxin. The link below describes the columns as well as distinguishes between when to use EndoTrap HD vs. Red.

https://ilexlife.com/products/endotrap-hd-endotoxin-removal-products

Brad