We are preparing a series of recombinant protein antigens and would appreciate recommendations for endotoxin removal before injections for antibody production.
Anion exchange is a very effective mode for removing Endotoxin and is routinely used for purification of biotherapeutics that are microbially-produced at the manufacturing scale. A simple DEAE or Q-Sepharose spin column should suffice due to the high concentration of negative charges associated with Endotoxin. Requires an extremely high concentration of salt to desorb from the solid phase.
Dr Palmieri is correct Q or DEAE sepharose is very effective. However, it also requires some knowledge of cation/anion purification and you may run the risk of binding your proteins by accident. If your group does not have expertise in sepharose purification I would highly recommend you just buy an endotoxin removal kit from Pierce thermo, Sigma, or Lonza. They are cheap, easy to use and highly specific to endotoxin. You also won't run the risk of accidently binding your proteins.
In my experience, the membrane chromatography Mustang E from PALL works quite well and are really easy to use. You may go for the Acrodiscs to try, they are siringe-filter sise membrane and come with instructios for a fast setup. Explore conditions in wich your interest protein does not bind to the mebrane. It works as an ion exchange chormatography
Astor Mignone I tried looking for the protocol for Acrodisc Mustang E on their website but I couldn't find it, Please I will like to know the procedure and what syringe size and buffer are needed for the process. Thanks
- Badges
- Science topic
- Similar topics
- Microbiology
- Assays