# US 2013/0153499 A1 · Separation medium for chromatography of various biomolecules > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2013/0153499 A1 - Document ID: US20130153499A1 - Status: Published application - Kind: A1 - Application number: 13/771,302 - Filed: 2013-02-20 - Published: 2013-06-20 - Earliest claimed: 2008-07-08 - Assignees: GE Healthcare Bio-Sciences AB - Inventors: GUNNAR GLAD; BO-LENNART JOHANSSON; JEAN-LUC MALOISEL; NILS NORRMAN - Web page: https://patentgrep.com/patent/US20130153499A1 ## CPC - B01D 15/3828: {Ligand exchange chromatography, e.g. complexation, chelation or metal interaction chromatography} - B01D 15/1896: {Membrane chromatography or membrane adsorbers} - B01D 15/36: involving ionic interaction, e.g. ion-exchange, ion-pair, ion-suppression or ion-exclusion - B01J 20/28092: {Bimodal, polymodal, different types of pores or different pore size distributions in different parts of the sorbent} - B01J 20/285: based on polymers - B01J 20/286: Phases chemically bonded to a substrate, e.g. to silica or to polymers - B01J 20/3242: {Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group} - B01J 20/3293: {Coatings on a core, the core being particle or fiber shaped, e.g. encapsulated particles, coated fibers} - C07K 1/16: by chromatography - C07K 1/22: Affinity chromatography or related techniques based upon selective absorption processes - C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies - B01D 15/166: {Fluid composition conditioning, e.g. gradient} - B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28) - B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract The present invention relates to a separation medium, comprising an inner core of a porous material provided with charged ligands, and an outer lid comprising a porous material provided with charged ligands, wherein the charge of the ligands in the inner core is opposite that of the charge of the ligands in the lid. The present invention also relates to a method for biomolecule separation comprising applying a sample to the above separation medium, wherein large molecules are prevented from entering the medium by charge repulsion from the medium and small molecules are captured in the inner core. ## Claims ### Claim at position 1 1-9. (canceled) ### Claim 10 A method for biomolecule separation comprising applying a sample to a separation medium, comprising an inner core of a porous material provided with charged ligands, and an outer lid comprising a porous material provided with charged ligands, wherein the charge of the ligands in the inner core is opposite that of the charge of the ligands in the lid, wherein large biomolecules are prevented from entering the separation medium by charge repulsion from the separation medium and small biomolecules are captured in said inner core. ### Claim 11 (depends on 10) The method of claim 10, wherein the charge repulsion is caused by the charged ligands in the lid. ### Claim 12 (depends on 11) The method of claim 11, wherein the charge repulsion, and thereby the prevention of entering of large biomolecules, is accompanied by a lower porosity (=steric hindrance) in the lid compared to the inner core. ### Claim 13 (depends on 10) The method of claim 10, wherein large biomolecules are obtained in the flow through by charge repulsion and small biomolecules adsorbed in the inner core are eluted from the separation medium. ### Claim 14 (depends on 13) The method of claim 13, wherein the small biomolecules are eluted with a salt gradient. ### Claim 15 (depends on 13) The method of claim 13, wherein the small biomolecules are eluted with a salt gradient and/or a pH gradient, in such a way that a portion of the small biomolecules gradually adhere to the ligands in the lid on their way out from the separation medium. ### Claim 16 (depends on 10) The method of claim 10, wherein the method is run in column format and the desired biomolecules are obtained in the flow-through. ### Claim 17 (depends on 10) The method of claim 10, wherein the method is run in column format and the desired biomolecules are obtained by elution from the inner core of the separation medium. ### Claim 18 (depends on 10) The method of claim 10, wherein the porous material in the inner core has the same porosity as the porous material in the lid. ### Claim 19 (depends on 10) The method of claim 10, wherein the porosity of the inner core and lid allows penetration of a biomolecule with a molecular weight of less than 100 000 g/mol. ### Claim 20 (depends on 19) The separation medium of claim 19, wherein the porosity of the inner core and lid allows penetration of a biomolecule with a molecular weight of less than 50 000 g/mol. ### Claim 21 (depends on 19) The separation medium of claim 19, wherein the amount of charged ligands is higher in the lid than in the core. ### Claim 22 (depends on 10) The method of claim 10, wherein the porous material in the inner core has the larger porosity than the porous material in the lid. ### Claim 23 The method of 22, wherein the porosity of the lid is such that biomolecules larger or equal to a molecular weight of 20 000, preferably 50 000 or 100 000 g/mol are excluded, and the porosity of the inner core is larger than the lid porosity.