# US 2010/0167374 A1 · Ultra-fast chromatography > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2010/0167374 A1 - Document ID: US20100167374A1 - Status: Published application - Kind: A1 - Application number: 12/528,670 - Filed: 2008-02-21 - Published: 2010-07-01 - Earliest claimed: 2007-02-26 - Assignees: Alltech Associates, Inc. - Inventors: Reno T. Nguyen; Scott Anderson; Ian Chappell - Web page: https://patentgrep.com/patent/US20100167374A1 ## CPC - C07K 1/22: Affinity chromatography or related techniques based upon selective absorption processes - B01D 15/325: {Reversed phase} - B01J 20/28004: {Sorbent size or size distribution, e.g. particle size} - B01J 20/28083: {being in the range 2-50 nm, i.e. mesopores} - B01J 20/283: based on silica - B01J 20/287: Non-polar phases; Reversed phases - C07K 1/20: Partition-, reverse-phase or hydrophobic interaction chromatography - C07K 16/065: {Purification, fragmentation} - 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) - 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 ## Abstract The present invention relates to a chromatographic method of separating biological material comprising, providing chromatographic media comprising inorganic oxide particles having an average diameter of about 2 microns or less and an average pore diameter of 300 Å or more; applying a solvent comprising said biological material to said media, wherein said biological material is reversibly bonded to said media; and eluting said biological material from said media with a solvent in less than about 2 minutes for biological material having a molecular weight of less than about 100,000 Daltons. ## Claims ### Claim 1 A chromatographic method of separating biological material comprising: providing chromatographic media comprising inorganic oxide particles having an average diameter of about 2 microns or less and an average pore diameter of 300 Å or more; applying a solvent comprising said biological material to said media, wherein said biological material is reversibly bonded to said media; and eluting said biological material from said media with a solvent in less than about 2 minutes for biological material having a molecular weight of less than about 100,000 Daltons. ### Claim 2 (depends on 1) A method according to claim 1, wherein said biological material comprises proteins, peptides. ### Claim 3 (depends on 1) A method according to claim 1, wherein said biological material comprises proteins. ### Claim 4 (depends on 3) A method according to claim 3, wherein said proteins comprise cytochrome c, ribonuclease, carbonic anhydrase II, bovine serum albumin, human serum albumin, human growth hormone, hemoglobin, IgG, conalbumin, ovalbumin, myoglobin, lysozyme, and insulin, or mixtures thereof. ### Claim 5 (depends on 1) A method according to claim 1, wherein said inorganic oxide particles comprise a median particle size ranging from about 1.0 μm to about 2.0 μm. ### Claim 6 (depends on 1) A method according to claim 1, wherein said solvent comprises water, acetonitrile, and trifluoroacetic acid. ### Claim 7 (depends on 1) A method according to claim 1, wherein said solvent comprises 0.08 to 0.1% v/v trifluoroacetic acid in water and acetonitrile. ### Claim 8 (depends on 1) A method according to claim 1, wherein said eluting of said biological material from said media comprises less than about 1.5 minutes. ### Claim 9 (depends on 1) A method according to claim 1, wherein said eluting of said biological material from said media comprises less than about 1.2 minutes. ### Claim 10 A chromatographic method of separating biological material comprising: providing chromatographic media comprising inorganic oxide particles having an average diameter of about 2 microns or less and an average pore diameter of 300 Å or more; applying a solvent comprising said biological material to said media, wherein said biological material is reversibly bonded to said media; and eluting said biological material from said media with a solvent in less than about 2 minutes with a chromatographic peak width of said eluted biological material being less than or equal to 0.1 minute. ### Claim 11 (depends on 10) A method according to claim 10, wherein said biological material comprises proteins, peptides. ### Claim 12 (depends on 10) A method according to claim 10, wherein said biological material comprises proteins. ### Claim 13 (depends on 12) A method according to claim 12, wherein said proteins comprise cytochrome c, ribonuclease, carbonic anhydrase II, bovine serum albumin, human serum albumin, human growth hormone, hemoglobin, IgG, conalbumin, ovalbumin, myoglobin, lysozyme, and insulin, or mixtures thereof. ### Claim 14 (depends on 10) A method according to claim 10, wherein said inorganic oxide particles comprise a median particle size ranging from about 1.0 μm to about 2.0 μm. ### Claim 15 (depends on 10) A method according to claim 10, wherein said solvent comprises water, acetonitrile, and trifluoroacetic acid. ### Claim 16 (depends on 10) A method according to claim 10, wherein said solvent comprises 0.08 to 0.1% v/v trifluoroacetic acid in water and acetonitrile. ### Claim 17 (depends on 10) A method according to claim 10, wherein said eluting of said biological material from said media comprises less than about 1.5 minutes. ### Claim 18 (depends on 10) A method according to claim 10, wherein said chromatograph peak width of said eluted biological material is less than about 0.07 minute. ### Claim 19 A chromatographic method of separating biological material comprising: providing chromatographic media comprising inorganic oxide particles having an average diameter of about 2 microns or less and an average pore diameter of 300 Å or more; applying a solvent comprising said biological material to said media, wherein said biological material is reversibly bonded to said media; and eluting said biological material from said media with a solvent in less than about 2 minutes while generating a backpressure of less than 3000 psi. ### Claim 20 (depends on 19) A method according to claim 19, wherein said biological material comprises proteins, peptides. ### Claim 21 (depends on 19) A method according to claim 19, wherein said biological material comprises proteins. ### Claim 22 (depends on 21) A method according to claim 21, wherein said proteins comprise cytochrome c, ribonuclease, carbonic anhydrase II, bovine serum albumin, human serum albumin, human growth hormone, hemoglobin, IgG, conalbumin, ovalbumin, myoglobin, lysozyme, and insulin, or mixtures thereof. ### Claim 23 (depends on 19) A method according to claim 19, wherein said inorganic oxide particles comprise a median particle size ranging from about 1.0 μm to about 2.0 μm. ### Claim 24 (depends on 19) A method according to claim 19, wherein said solvent comprises water, acetonitrile, and trifluoroacetic acid. ### Claim 25 (depends on 19) A method according to claim 19, wherein said solvent comprises 0.08 to 0.1% v/v trifluoroacetic acid in water and acetonitrile. ### Claim 26 (depends on 19) A method according to claim 19, wherein said eluting of said biological material from said media comprises less than about 1.5 minutes. ### Claim 27 (depends on 19) A method according to claim 19, wherein said backpressure is less than about 2800 psi. ### Claim 28 A chromatographic method of separating at least one antibody comprising: providing chromatographic media comprising inorganic oxide particles; applying a solvent comprising said antibody to said media, wherein said antibody is reversibly bonded to said media; and eluting said antibody from said media with a solvent in less than about 4 minutes, wherein said antibody remains intact after elution. ### Claim 29 (depends on 28) A method according to claim 28, wherein said antibody comprises IgG. ### Claim 30 (depends on 28) A method according to claim 28, wherein said antibody comprises human or animal IgG. ### Claim 31 (depends on 28) A method according to claim 28, wherein said inorganic oxide particles comprise a median particle size ranging from about 1.0 μm to about 2.0 μm. ### Claim 32 (depends on 28) A method according to claim 28, wherein said solvent comprises water, n-propanol, and trifluoroacetic acid. ### Claim 33 (depends on 28) A method according to claim 28, wherein said solvent comprises 0.8 to 0.1% v/v trifluoroacetic acid in water and n-propanol. ### Claim 34 (depends on 28) A method according to claim 28, wherein said eluting of said antibody from said media comprises less than about 3.5 minutes. ### Claim 35 (depends on 28) A method according to claim 28, wherein 100% by weight of said antibody remains intact after said eluting. ### Claim 36 A chromatographic method of separating at least one antibody comprising: providing chromatographic media comprising inorganic oxide particles; applying a solvent comprising said antibody to said media, wherein said antibody is reversibly bonded to said media; and eluting said antibody from said media with a solvent in less than about 4 minutes with a chromatographic peak width of said eluted antibody being less than or equal to about 0.5 minute. ### Claim 37 (depends on 36) A method according to claim 36, wherein said antibody comprises IgG. ### Claim 38 (depends on 36) A method according to claim 36, wherein said antibody comprises human or animal IgG. ### Claim 39 (depends on 36) A method according to claim 36, wherein said inorganic oxide particles comprise a median particle size ranging from about 1.0 μm to about 2.0 μm. ### Claim 40 (depends on 36) A method according to claim 36, wherein said solvent comprises water, n-propanol, and trifluoroacetic acid. ### Claim 41 (depends on 36) A method according to claim 36, wherein said solvent comprises 0.8 to 0.1% v/v trifluoroacetic acid in water and n-propanol. ### Claim 42 (depends on 36) A method according to claim 36, wherein said eluting of said antibody from said media comprises less than about 3.5 minutes. ### Claim 43 (depends on 36) A method according to claim 36, wherein said chromatographic peak width of said eluted biological material is less than or equal to 0.4 minute.