# US 11,079,361 B2 · Chromatography column qualification in manufacturing methods for producing anti-IL12/IL23 antibody compositions > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,079,361 B2 - Document ID: US11079361B2 - Status: Granted - Kind: B2 - Application number: 16/389,114 - Filed: 2019-04-19 - Published: 2019-10-24 - Granted: 2021-08-03 - Earliest claimed: 2018-04-20 - Assignees: Janssen Biotech, Inc. - Applicants: Janssen Biotech, Inc. - Inventors: Paul Randolph - Web page: https://patentgrep.com/patent/US11079361B2 ## CPC - C07K 16/244: {Interleukins [IL]} - G01N 30/88: Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 - A61K 39/39591: {Stabilisation, fragmentation} - B01D 15/166: {Fluid composition conditioning, e.g. gradient} - B01D 15/20: relating to the conditioning of the sorbent material - B01D 15/206: {Packing or coating} - B01D 15/362: {Cation-exchange} - B01D 15/363: {Anion-exchange} - B01D 15/3809: {of the antigen-antibody type, e.g. protein A, G or L chromatography} - C07K 1/18: Ion-exchange chromatography - C07K 1/22: Affinity chromatography or related techniques based upon selective absorption processes - C07K 16/065: {Purification, fragmentation} - C07K 16/241: {Tumor Necrosis Factors} - G01N 30/56: Packing methods or coating methods - G01N 30/86: Signal analysis - G01N 30/8665: {for calibrating the measuring apparatus} - G01N 30/89: Inverse chromatography - G01N 35/00623: {of instruments} - C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered - C07K 2317/31: multispecific - C07K 2317/565: Complementarity determining region [CDR] - G01N 2030/562: {packing} - G01N 2030/8831: {involving peptides or proteins} - G01N 2030/889: {monitoring the quality of the stationary phase; column performance} - 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) - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - 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) ## Abstract A method of operating a chromatography column is described for use in methods of manufacture for producing anti-IL-12/IL-23p40 antibodies, e.g., the anti-IL-12/IL-23p40 antibody STELARA® (ustekinumab). This method involves collecting column outlet signal and accumulated flow parameters at two or more intervals of at least one mobile phase transition front during operation of the chromatography column comprising column packing. A model gamma cumulative distribution curve is calculated based on the collected column outlet signal and accumulated flow parameters for the at least one mobile phase transition front. A height equivalent theoretical plate (HETP) value is calculated for the at least one mobile phase transition front using parameters of the model gamma cumulative distribution curve and the quality of the chromatography column packing is assessed based on the calculated HETP value. ## Claims ### Claim 1 A method of operating a chromatography column in methods of manufacture for producing anti-IL-12/IL-23p40 antibodies, specific pharmaceutical compositions of the antibodies, and antigen binding fragments thereof, wherein the anti-IL-12/IL-23p40 antibodies comprise amino acid sequences selected from the group consisting of: (i) a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO:10 and a light chain (LC) comprising the amino acid sequence of SEQ ID NO:11; (ii) a heavy chain variable domain amino acid sequence of SEQ ID NO:7 and a light chain variable domain amino acid sequence of SEQ ID NO:8; and (iii) heavy chain CDR amino acid sequences of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3, and light chain CDR amino acid sequences of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6, said method comprising: collecting a column outlet signal and accumulated flow parameters at two or more intervals of at least one mobile phase transition front during a first operation of the chromatography column comprising column packing; determining a model gamma cumulative distribution curve based on the collected column outlet signal and accumulated flow parameters for the at least one mobile phase transition front using Formula Ia for a rising transition front or Formula Ib for a falling transition front, C = 1 - 1 r ⁡ ( k ) ⁢ γ ⁡ ( k , V - V i θ ) Formula ⁢ ⁢ Ia or C = 1 r ⁡ ( k ) ⁢ γ ⁡ ( k , V - V i θ ) Formula ⁢ ⁢ Ib wherein C is the column outlet signal for a given V, V is the accumulated flow divided by column volume, and k, θ, and Vi are the shape, scale and offset parameters, respectively, used to define the curve; calculating a height equivalent theoretical plate (HETP) value for the at least one mobile phase transition front using Formula II and the model gamma cumulative distribution curve parameters of k, θ, and Vi, HETP = σ 2 μ 2 ⁢ L Formula ⁢ ⁢ II wherein μ=kθ+Vi σ=√{square root over (kθ2)} L=column length; and assessing the quality of the chromatography column packing based on said calculated HETP value. ### Claim 2 (depends on 1) The method of claim 1, further comprising: conditioning, replacing, or repacking the chromatography column based on said assessing. ### Claim 3 (depends on 2) The method of claim 2 further comprising: collecting column outlet signals and accumulated flow parameters at two or more intervals of a corresponding mobile phase transition front during one or more subsequent uses of the chromatography column packing; performing said determining and said calculating using the column outlet signals and accumulated flow parameters collected during each of the one or more subsequent uses of the chromatography column packing; determining an HETP value of the chromatography column packing during each of said one or more subsequent uses based on said performing; compiling a trend of the determined HETP values of the chromatography column packing of the one or more subsequent uses; and identifying a change in the quality of the chromatography column packing based on said compiled trend, wherein said conditioning, replacing or repacking of the chromatography column is based on said identifying. ### Claim 4 (depends on 3) The method of claim 3, wherein an increase in the HETP value of the chromatography column packing in the one or more subsequent uses of said column packing as compared to the HETP value of the chromatography column packing in one or more earlier uses of said column packing identifies a decrease in the quality of the chromatography column packing. ### Claim 5 (depends on 2) The method of claim 2, wherein column outlet signals and accumulated flow parameters of two or more different mobile phase transition fronts during said first operation of the column packing are collected, said method comprising: performing said determining and calculating using the column outlet signals and accumulated flow parameters collected for each of the two or more different mobile phase transition fronts independently to calculate an HETP value for each of the two of more different mobile phase transition fronts; assessing the quality of the chromatography column packing based on the two or more calculated HETP values, whereby said conditioning, replacing or repacking the chromatography column is based on said assessing. ### Claim 6 (depends on 1) The method of claim 1, wherein the chromatography column is selected from the group consisting of: a Protein A affinity chromatography column, a cation exchange chromatography column, and an anion exchange chromatography column. ### Claim 7 (depends on 6) The method of claim 6, wherein the Protein A affinity chromatography column comprises a MABSELECT® Protein A affinity chromatography column, the cation exchange chromatography column comprises a UNOSPHERE™ S cation exchange chromatography column or an SP SEPHAROSE® XL cation exchange chromatography column, and the anion exchange chromatography column comprises a Q SEPHAROSE® XL anion exchange chromatography column. ### Claim 8 (depends on 6) The method of claim 6, wherein the mobile phase transition front in the Protein A affinity chromatography column is generated from one or more fronts selected from the group consisting of: a wash front generated during purification of the anti-IL-12/IL-23p40 antibodies, a front generated during elution of the anti-IL-12/IL-23p40 antibodies, a front generated during sanitization of the column with guanidine HCl, and a front generated during post-sanitization rinsing of the column with 0.1 M sodium citrate, pH 3.5. ### Claim 9 (depends on 6) The method of claim 6, wherein the mobile phase transition front in the cation exchange chromatography column is generated from one or more fronts selected from the group consisting of: a front generated during loading of solvent/detergent (S/D) treated material comprising the anti-IL-12/IL-23p40 antibodies, a front generated during elution of the anti-IL-12/IL-23p40 antibodies, and a front generated during a column strip. ### Claim 10 (depends on 6) The method of claim 6, wherein the mobile phase transition front in the anion exchange chromatography column is generated from one or more fronts selected from the group consisting of: a front generated during cleaning of the column with sodium hydroxide and a front generated during a column strip. ### Claim 11 (depends on 3) The method of claim 3, wherein the chromatography column is selected from the group consisting of: a Protein A affinity chromatography column, a cation exchange chromatography column, and an anion exchange chromatography column. ### Claim 12 (depends on 11) The method of claim 11, wherein the Protein A affinity chromatography column comprises a MABSELECT® Protein A affinity chromatography column, the cation exchange chromatography column comprises a UNOSPHERE™ S cation exchange chromatography column, and the anion exchange chromatography column comprises a Q SEPHAROSE® XL anion exchange chromatography column. ### Claim 13 (depends on 11) The method of claim 11, wherein the mobile phase transition front in the Protein A affinity chromatography column is generated from one or more fronts selected from the group consisting of: a front generated during elution of anti-TNF antibodies, a front generated during sanitization of the column with guanidine HCl, and a front generated during post-sanitization rinsing of the column with 0.1 M sodium citrate, pH 3.5. ### Claim 14 (depends on 11) The method of claim 11, wherein the mobile phase transition front in the cation exchange chromatography column is generated from one or more fronts selected from the group consisting of: a front generated during loading of solvent/detergent (S/D) treated material comprising anti-TNF antibodies, a front generated during elution of the anti-TNF antibodies, and a front generated during a column strip. ### Claim 15 (depends on 11) The method of claim 11, wherein the mobile phase transition front in the anion exchange chromatography column is generated from one or more fronts selected from the group consisting of: a front generated during cleaning of the column with sodium hydroxide and a front generated during a column strip. ### Claim 16 (depends on 5) The method of claim 5, wherein the chromatography column is selected from the group consisting of: a Protein A affinity chromatography column, a cation exchange chromatography column, and an anion exchange chromatography column. ### Claim 17 (depends on 16) The method of claim 16, wherein the Protein A affinity chromatography column comprises a MABSELECT® Protein A affinity chromatography column, the cation exchange chromatography column comprises a UNOSPHERE™ S cation exchange chromatography column, and the anion exchange chromatography column comprises a Q SEPHAROSE® XL anion exchange chromatography column. ### Claim 18 (depends on 16) The method of claim 16, wherein the mobile phase transition front in the Protein A affinity chromatography column is generated from one or more fronts selected from the group consisting of: a front generated during elution of anti-TNF antibodies, a front generated during sanitization of the column with guanidine HCl, and a front generated during post-sanitization rinsing of the column with 0.1 M sodium citrate, pH 3.5. ### Claim 19 (depends on 16) The method of claim 16, wherein the mobile phase transition front in the cation exchange chromatography column is generated from one or more fronts selected from the group consisting of: a front generated during loading of solvent/detergent (S/D) treated material comprising anti-TNF antibodies, a front generated during elution of the anti-TNF antibodies, and a front generated during a column strip. ### Claim 20 (depends on 16) The method of claim 16, wherein the mobile phase transition front in the anion exchange chromatography column is generated from one or more fronts selected from the group consisting of: a front generated during cleaning of the column with sodium hydroxide and a front generated during a column strip.