Method for ion-exchange chromatography and media used thereof
View Patent ↗The present invention relates to a method for running ion exchange chromatography on a media comprising shell beads having an inner porous core and an outer shell, wherein the inner core is provided with ligands whose charge changes with pH and the shell is provided with charged ion exchange ligands, the method comprising the following steps: a) adsorbing sample molecules on the shell ligands at a first pH; b) causing a discharge of the inner core ligands at a second pH by addition of a buffer substance that is able to increase its charge having the same sign/type as that of the core ligands, which at the same time causes release of ions from the inner core ligands and thereby an increase in ionic strength that displaces the sample molecules from the shell ligands i.e. causes an elution.
1. A method for running ion exchange chromatography, the method comprising the following steps:
a) providing an ion exchange chromatographic media comprising shell beads having an outer shell and an inner porous core, wherein said outer shell has strong ion exchange ligands and said inner core is functionalized with weak ion exchange ligands;
b) injecting a sample containing charged molecules onto said media thereby causing the sample molecules to be absorbed on the outer shell ligands at a first pH;
c) modifying the pH of the sample by applying a buffer having a second pH, thereby causing a decrease in charge in said inner core ligands and a release of ions that displaces the sample molecules from the outer shell ligands and causes the sample molecules to be eluted from the media.
2. The method of claim 1 , wherein the strong ion exchange ligands comprise —N 30 (CH 3 ) 3 and all types of quaternary ammonium ligands and non aromatic and aromatic —SO 3 − ligands.
3. The method of claim 1 , wherein the charge change per pH unit of functionalized core is >30 μmol/pH·mL media.
4. The method of claim 1 , wherein the outer shell strong ion exchange ligands and said inner core weak ion exchange ligands have the same or different charge and are selected from cation ligands and anion ligands.
5. The method of claim 1 , wherein the porosity of said inner core prevents sample molecules from penetrating into said inner core.
6. The method of claim 1 , wherein the porosity of said inner core allows sample molecules to penetrate into said inner core.
7. The method of claim 1 , wherein the shell beads have a diameter between 3 to 400 μm.
8. The method of claim 7 , wherein the outer shell is 0.5-100 μm thick.
9. The method of claim 1 , wherein the method is run in column format and the direction of flow is reversed in the elution step.
10. A chromatography media comprising shell beads having an inner porous core and an outer shell, wherein said outer shell has strong ion exchange ligands and said inner core is functionalized with weak ion exchange ligands wherein the density of the weak ion exchange ligands is effective to cause charge change per pH unit >30 μmol/pH·mL media.