Method for endotoxin removal
View Patent ↗The present invention relates to a method for endotoxin removal from a sample comprising the following steps: combining the sample comprising one or more target molecule(s) with a chromatography media comprising beads having an inner porous core functionalized with ligands capable of binding endotoxin and an outer porous layer without functional groups and a pore size small enough to exclude the target molecule from the inner core; and collecting the sample from the media, wherein the sample comprises an endotoxin level which is at least 75% less, preferably 90% less, than before the removal and the yield of the target molecule is at least 75%.
1. A method for endotoxin removal from a sample comprising the following steps:
combining the sample comprising one or more target molecule(s) and an endotoxin with a chromatography media comprising beads having an inner porous core functionalized with ligands capable of binding the endotoxin and a 5-7 μm thick outer porous layer without functional groups and a pore size small enough to exclude the target molecule from the inner core, wherein the exclusion limit of said outer porous layer is about 700 kDa; and
collecting the sample from the media, wherein the sample comprises an endotoxin level which is at least 75% less, preferably 90% less, than before the removal and the yield of the target molecule is at least 75%.
2. Method according to claim 1 , wherein the chromatography media is packed in a column and the sample is collected in the flow-through.
3. The method of claim 1 , wherein the target molecule is a virus.
4. The method of claim 1 , wherein the core is functionalized with ligands capable of binding endotoxins such as primary, secondary, tertiary or quaternary amines.
5. The method of claim 4 , wherein the ligands are multimodal amines containing a hydrophobic moiety.
6. The method of claim 5 , wherein the hydrophobic moiety consists of an aliphatic or aromatic hydrocarbon with 3-20 carbon atoms, preferably between 4 and 18 carbon atoms.
7. The method of claim 6 , wherein the ligand is octylamine.
8. The method of claim 1 , wherein the chromatography media is made of a natural or synthetic polymer.
9. The method of claim 1 , wherein the sample is a supernatant from a cell culture producing recombinant protein or virus.
10. The method of claim 1 , wherein the chromatography media is made of a natural polymer.
11. The method of claim 1 , wherein the chromatography media is made of agarose.