Granted Patent
B2
US 12,595,294 · App. 17/812,145 · Granted Apr 7, 2026
Methods to decrease impurities from recombinant protein manufacturing processes
Assignee:
Hoffmann-La Roche Inc.
View Patent ↗
Loading inventors, assignments & file history…
Abstract
The invention relates to a method of reducing the amount of non-aggregate produce-related impurities (NAPRIs) in a buffered solution of monoclonal antibodies (mAbs), involving the use of a synthetic depth filter. The invention may be of use in the purification of monoclonal 5 antibodies.
Claims (19)
1 . A method of producing a buffered solution of monoclonal antibodies (mAbs) with a reduced amount of non-aggregate product-related impurities (NAPRIs), the method comprising passing an initial buffered solution comprising: (a) mAbs, and (b) NAPRIs, through a synthetic depth filter comprising silica and polyacrylic fiber to produce the buffered solution of mAbs with the reduced amount of NAPRIs,
wherein the NAPRIs comprise incompletely or incorrectly assembled mAb polypeptides,
wherein the NAPRIs comprise high molecular weight (HMW) polypeptides having a molecular weight higher than the mAb,
wherein the NAPRIs do not comprise aggregates, and
wherein the reduced amount of NAPRIs is with respect to the amount of NAPRIs in the initial buffered solution before it has passed through the synthetic depth filter comprising silica and polyacrylic fiber.
2 . The method of claim 1 , wherein the mAb is a multispecific antibody.
3 . The method of claim 1 , wherein the mAb is an antibody fusion protein comprising an antibody or antibody fragment and another biologically active polypeptide.
4 . The method of claim 1 , wherein the initial buffered solution of mAbs is passed through the depth filter at a temperature that is between 4° C. and 22° C.
5 . The method of claim 1 , wherein the NAPRI is a polypeptide lacking one or more polypeptide chains of the mAb and/or the NAPRI is a polypeptide comprising a different polypeptide chain arrangement than the mAb.
6 . The method of claim 1 , wherein the NAPRI comprises two heavy chains having the same amino acid sequence.
7 . The method of claim 1 , wherein the depth filter is a dual layer depth filter.
8 . The method of claim 1 , wherein the depth filter does not contain diatomaceous earth.
9 . The method of claim 1 , further comprising measuring the NAPRI concentration in the buffered solution of mAbs, wherein the NAPRI concentration is measured by Size Exclusion Chromatography (SEC) and/or with Capillary Electrophoresis SDS Page in a non-reducing environment.
10 . The method of claim 1 , wherein the initial buffered solution of mAbs has been subjected to chromatography before passing through the synthetic depth filter, and wherein the chromatography comprises one or more of ion exchange chromatography, anion exchange column, a cation exchange column, or multimodal (mixed mode) chromatography.
11 . The method of claim 1 , wherein the initial buffered solution of mAbs is passed through the depth filter at a temperature of between 10° C. and 21° C., or between 15° C. and 20° C.
12 . The method of claim 1 , wherein the initial buffered solution of mAbs is passed through the depth filter at mass load of in the range of 100 g/m 2 to 2500 g/m 2 , 300 g/m 2 to 2000 g/m 2 , or 500 g/m 2 to 1500 g/m 2 .
13 . The method of claim 1 , wherein the initial buffered solution of mAbs has a pH in the range of 4.0 to 7.5, 4.0 to 7.2, or 4.0 to 5.5 when it passes through the depth filter.
14 . The method of claim 1 , wherein the initial buffered solution of mAbs is passed through the depth filter at a flow rate in the range of 1 L/min*m 2 to 10 L/min*m 2 , 1.5 L/min*m 2 to 8 L/min*m 2 , or at a flow rate that is 4.3 L/min*m 2 .
15 . The method of claim 1 , further comprising the step of isolating the mAbs from the buffered solution of mAbs.
Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Apr 7, 2023
From: FEISTL, CHRISTOPH; POMPIATI, MARC
To: ROCHE DIAGNOSTICS GMBH
Reel/Frame 063262/0471 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Apr 7, 2023
From: ROCHE DIAGNOSTICS GMBH
To: F. HOFFMANN-LA ROCHE AG
Reel/Frame 063262/0502 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Apr 7, 2023
From: F. HOFFMANN-LA ROCHE AG
To: HOFFMANN-LA ROCHE INC.
Reel/Frame 063262/0518 →
Continuity (2)
Continuation
PCTEP2021050809
· Jan 15, 2021
References Cited (143)
US 4676980A
· Segal
· 1987
[cited by applicant]
US 4882013A
· Turner et al.
· 1989
[cited by applicant]
US 5591828A
· Bosslet
· 1997
[cited by applicant]
US 5731168A
· Carter
· 1998
[cited by applicant]
US 9249182B2
· Herigstad et al.
· 2016
[cited by applicant]
US 20140010820A1
· Wang et al.
· 2014
[cited by applicant]
US 20160176921A1
· Rajendran et al.
· 2016
[cited by applicant]
US 20160272674A1
· Althouse et al.
· 2016
[cited by applicant]
US 20170073396A1
· Bataille et al.
· 2017
[cited by applicant]
US 20180360856A1
· Holmes et al.
· 2018
[cited by applicant]
US 20180369258A1
· Holmes et al.
· 2018
[cited by applicant]
US 20220135620A1
· Seay et al.
· 2022
[cited by applicant]
EP 0404097A2
· 1990
[cited by applicant]
WO 199311161A1
· 1993
[cited by applicant]
WO 199850431A2
· 1998
[cited by applicant]
WO 199850431A3
· 1999
[cited by applicant]
WO 200177342A1
· 2001
[cited by applicant]
WO 2003100080A1
· 2003
[cited by applicant]
WO 2006044532A1
· 2006
[cited by applicant]
WO 2008024715A2
· 2008
[cited by applicant]
WO 2008024715A3
· 2008
[cited by applicant]
WO 2009080251A1
· 2009
[cited by applicant]
WO 2009080252A1
· 2009
[cited by applicant]
WO 2009080253A1
· 2009
[cited by applicant]
WO 2009089004A1
· 2009
[cited by applicant]
WO 2010048192A2
· 2010
[cited by applicant]
WO 2010048192A3
· 2010
[cited by applicant]
WO 2010112193A1
· 2010
[cited by applicant]
WO 2010115589A1
· 2010
[cited by applicant]
WO 2010136172A1
· 2010
[cited by applicant]
WO 2010145792A1
· 2010
[cited by applicant]
WO 2011034605A2
· 2011
[cited by applicant]
WO 2011034605A3
· 2011
[cited by applicant]
WO 2011150110A1
· 2011
[cited by applicant]
WO 2013009491A2
· 2013
[cited by applicant]
WO 2013026831A1
· 2013
[cited by applicant]
WO 2013028330A2
· 2013
[cited by applicant]
WO 2013028330A3
· 2013
[cited by applicant]
WO 2013177115A2
· 2013
[cited by applicant]
WO 2014004281A1
· 2014
[cited by applicant]
WO 2013177115A3
· 2014
[cited by applicant]
WO 2015023468A1
· 2015
[cited by applicant]
WO 2015031899A1
· 2015
[cited by applicant]
WO 2015077605A1
· 2015
[cited by applicant]
WO 2015095539A1
· 2015
[cited by applicant]
WO 2015150447A1
· 2015
[cited by applicant]
WO 2015198320A1
· 2015
[cited by applicant]
WO 2016016299A1
· 2016
[cited by applicant]
WO 2016106291A1
· 2016
[cited by applicant]
WO 2016172485A2
· 2016
[cited by applicant]
WO 2016172485A3
· 2016
[cited by applicant]
WO 2017031476A2
· 2017
[cited by applicant]
WO 2017027861A1
· 2017
[cited by applicant]
WO 2017095062A1
· 2017
[cited by applicant]
WO 2017218977A2
· 2017
[cited by applicant]
WO 2017218977A3
· 2018
[cited by applicant]
WO 2018035025A1
· 2018
[cited by applicant]
WO 2018170488A1
· 2018
[cited by applicant]
WO 2018200430A1
· 2018
[cited by applicant]
WO 2019191416A1
· 2019
[cited by applicant]
WO 2020006266A1
· 2020
[cited by applicant]
WO 2020023566A1
· 2020
[cited by applicant]
WO 2020159838A1
· 2020
[cited by applicant]
WO 2020200980A1
· 2020
[cited by applicant]
WO 2020227144A1
· 2020
[cited by applicant]
WO 2021144422A1
· 2021
[cited by applicant]
WO 2022094116A1
· 2022
[cited by applicant]
Al-Lazikani, B. et al. (1997). “Standard Conformations for the Canonical Structures of Immunoglobulins,” J. Mol. Biol. 273:927-948.
[cited by applicant]
Atwell, S. et al. (1997). “Stable Heterodimers From Remodeling The Domain Interface Of A Homodimer Using A Phage Display Library,” J. Mol. Biol. 270 (1):26-35.
[cited by applicant]
Brennan, M. et al. (Jul. 5, 1985). “Preparation of Bispecific Antibodies by Chemical Recombination of Monoclonal Immunoglobulin G1 Fragments,” Science 229:81-83.
[cited by applicant]
Chadd, H.E. et al. (2001). “Therapeutic Antibody Expression Technology,” Curr. Opin. Biotechnol 12:188-194.
[cited by applicant]
Charlton, H.R. (Jan. 1, 1999). “Characterisation of a Generic Monoclorial Antibody Harvesting System For Adsorption of DNA By Depth Filters and Various Membranes,” Bioseparation 8:281-291, 27 pages.
[cited by applicant]
Chiu, J. et al. (May 2017). “Knockout of a Difficult-To-Remove CHO Host Cell Protein, Lipoprotein Lipase, For Improved Polysorbate Stability In Monoclonal Antibody Formulations,” Biotechnology And Bioengineering 114(5):…
[cited by applicant]
Chothia, C. et al. (1987). “Canonical Structures for the Hypervariable Regions of Immunoglobulins,” J. Mol. Biol. 196:901-917.
[cited by applicant]
Chothia, C. et al. (Dec. 21/28, 1989). “Conformations of Immunoglobulin Hypervariable Regions,” Nature 342(6252):877-883.
[cited by applicant]
Chothia, C. et al. (Dec. 5, 1985). “Domain Association In Immunoglobulin Molecules. The Packing Of Variable Domains,” J. Mol. Biol. 186(3):651-663.
[cited by applicant]
Extended European Search Report, dated Jun. 25, 2022, for European Patent Application No. 20151994.9, 9 pages.
[cited by applicant]
Giese, G. et al. (2018, e-pub. Nov. 29, 2017). “Bispecific Antibody Process Development: Assembly and Purification of Knob and Hole Bispecific Antibodies,” Biotechnol. Prog. 34(2):397-404. Abstract.
[cited by applicant]
Gruber, M. et al. (1994). “Efficient Tumor Cell Lysis Mediated by a Bispecific Single Chain Antibody Expressed in
[cited by applicant]
Hall, T. et al. (May 2006, e-pub. Apr. 5, 2016). “Polysorbates 20 and 80 Degradation by Group XV Lysosomal Phospholipase A2 Isomer XI in, Monoclonal Antibody Formulations,” Journal Of Pharmaceutical Sciences 105(5):1633…
[cited by applicant]
Holliger, P. et al. (Sep. 2005) “Engineered Antibody Fragments and The Rise Of Single Domains,” Nat. Biotechnol. 23(9):1126-1136.
[cited by applicant]
Hollinger, P. et al. (Jul. 1993). “Diabodies”: Small Bivalent And Bispecific Antibody Fragments, Proc. Natl. Acad. Sci. USA 90:6444-6448.
[cited by applicant]
International Preliminary Report on Patentability, issued Jul. 19, 2022, for PCT Application No. PCT/EP2021/050809, filed Jan. 15, 2021, 8 pages.
[cited by applicant]
International Preliminary Report on Patentability, issued Nov. 2, 2021, for PCT Application No. PCT/US2020/031164, filed May 1, 2020, 9 pages.
[cited by applicant]
International Search Report and Written Opinion, mailed Jul. 17, 2020, for PCT Application No. PCT/US2020/031164, filed May 1, 2020, 17 pages.
[cited by applicant]
International Search Report and Written Opinion, mailed Mar. 23, 2021, for PCT Application No. PCT/EP2021/050809, filed Jan. 15, 2021, 11 pages.
[cited by applicant]
International Search Report and Written Opinion, mailed Mar. 31, 2022 for PCT Application No. PCT/US2021/057100, filed Oct. 28, 2021, 24 pages.
[cited by applicant]
Invitation To Pay Additional Fees, mailed Feb. 10, 2022, for PCT Application No. PCT/US2021/057100, filed Oct. 28, 2021, 21 pages.
[cited by applicant]
Jones, P.T. et al. (May 29, 1986). “Replacing the Complementarity-Determining Regions in a Human Antibody With Those From a Mouse,” Nature 321:522-525.
[cited by applicant]
Kabat, E.A. et al. (1991). Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD. TOC, 21 pages.
[cited by applicant]
Klein, C. et al. (Jun. 10, 2016, e-pub. Jul. 11, 2016). “The Use of CrossMab Technology for the Generation of Bi- and Multispecific Antibodies,” MABS 8(6):1010-1020.
[cited by applicant]
Kostelny, S.A. et al. (Mar. 1, 1992). “Formation of a Bispecific Antibody By The Use of Leucine Zippers,” J. Immunol. 148(5):1547-1553.
[cited by applicant]
Liu, H. et al. (Sep./Oct. 2010). “Recovery and Purification Process Development for Monoclonal Antibody Production,” mAbs 2(5): 480-499.
[cited by applicant]
Marichal-Gallardo, P.A. et al. (2012, e-pub. Jun. 26, 2012). “State-Of-The-Art In Downstream Processing Of Monoclonal Antibodies: Process Trends In Design and Validation,” Biotechnology Progress 28(4):899-916.
[cited by applicant]
Merchant, A. M. et al. (Jul. 1998). “An Efficient Route To Human Bispecific IgG,” Nature Biotechnology 16:677-681.
[cited by applicant]
Milstein, C. et al. (Oct. 6, 1983). “Hybrid Hybridomas and Their Use In Immunohistochemistry,” Nature 305:537-540.
[cited by applicant]
Morrison, S.C. et al. (Nov. 1984). “Chimeric Human Antibody Molecules: Mouse Antigen-Binding Domains With Human Constant Region Domains,” Proc. Natl. Acad. Sci. USA 81:6851-6855.
[cited by applicant]
Nguyen, H.C. et al. (May 11, 2018). “Improved HCP Reduction Using a New, All-Synthetic Depth Filtration Media Within an Antibody Purification Process,” Biotechnology Journal 14(11):1700771, 11 pages.
[cited by applicant]
Onur, A. et al. (Sep. 12, 2018). “Multi-Layer Filters: Adsorption and Filtration Mechanisms for Improved Separation,” Frontiers in Chemistry 6(417):1-11.
[cited by applicant]
Plückthun, A. (1994). “Antibodies from
[cited by applicant]
Presta, L.G. (1992). “Antibody Engineering,” Current Opinion in Structural Biology, 2:593-596.
[cited by applicant]
Ridgway, J.B.B. et al. (1996). “‘Knobs-Into-Holes’ Engineering of Antibody CH3 Domains for Heavy Chain Heterodimerization,” Protein Engineering 9(7):617-621.
[cited by applicant]
Riechmann, L. et al. (Mar. 24, 1988). “Reshaping Human Antibodies for Therapy,” Nature 332:323-327.
[cited by applicant]
Schaefer, W. et al. (Jul. 5, 2011, e-pub. Jun. 20, 2011). “Immunoglobulin Domain Crossover as a Generic Approach for the Production of Bispecific IgG Antibodies,” Proc. Natl. Acad. Sci. U.S.A. 108(27):11187-11192.
[cited by applicant]
Singh, N. et al. (2017, e-pub. Jan. 12, 2017). “Development of Adsorptive Hybrid Filters to Enable Two-Step Purification of Biologics,” MABS 9(2):350-364.
[cited by applicant]
Spiess, C. et al. (2015, e-pub. Jan. 27, 2015). “Alternative Molecular Formats and Therapeutic Applications For Bispecific Antibodies,” Mol. Immunol. 67:95-106.
[cited by applicant]
Tutt, A. et al. (Jul. 1, 1991) “Trispecific F(ab')3 Derivatives that use Cooperative Signaling Via the TCR/CD3 Complex and CD2 to Activate and Redirect Resting Cytotoxic T Cells,” J. Immunol. 147(1):60-69.
[cited by applicant]
Van Dijk, M.A. et al. (Aug. 2001). “Human Antibodies as Next Generation Therapeutics,” Curr. Opin. Che. Biology 5(4):368-374.
[cited by applicant]
Yigzaw, Y. et al. (2006, e-pub. Jan. 1, 2006). “Exploitation of The Adsorptive Properties of Depth, Filters For Host Cell Protein Removal During Monoclonal Antibody Purification,” Biotechnology Progress 22(1):288-296.
[cited by applicant]
Yu, D. et al. (Jun. 11, 2019). “Control of Antibody High And Low Molecular Weight Species By Depth Filtration-Based Cell Culture Harvesting,” Biotechnology And Bioengineering 116(10):2610-2620.
[cited by applicant]
Zhou, J.X. et al. (Oct. 1, 2008). “Implementation of Advanced Technologies In Commercial Monoclonal Antibody Production,” Biotechnology Journal 3(9-10):1185-1200.
[cited by applicant]
U.S. Appl. No. 17/767,842, Davies et al, filed Apr. 8, 2022 (not submitted herewith pursuant to the waiver of 37 C.F.R. § 1.98(a)(2)(iii) issued by the Office on Sep. 21, 2004).
[cited by applicant]
U.S. Appl. No. 17/797,293, Arcadu et al, filed Aug. 3, 2022 (not submitted herewith pursuant to the waiver of 37 C.F.R. § 1.98(a)(2)(iii) issued by the Office on Sep. 21, 2004).
[cited by applicant]
International Preliminary Report on Patentability, issued May 2, 2023, for PCT Application No. PCT/US2021/057100, filed Oct. 28, 2021, 14 pages.
[cited by applicant]
3M Purification Inc. (Oct. 22, 2020). “Polisher ST Scale-up Capsules: Scale-Up Capsules Installation and Operation Instruction,” 34-8726-1126-3, Datasheet, 82 pages.
[cited by applicant]
3M Purification Inc. (Sep. 2018). “Safety Information & Installation and Operation Instructions,” 34-8723-4281-0 Datasheet, 80 pages.
[cited by applicant]
Amara, J. et al. (Sep. 2016). “Novel Synthetic Adsorptive Depth Filter Media for CHO Harvest Clarification,” Merck Poster, 1 page.
[cited by applicant]
Anonymous (Jun. 2018). MILLSTAK+® HC Pro Data Sheet, Merck, 5 pages.
[cited by applicant]
Anonymous (Jun. 2020). “Prefilter Selection Guide,” Merck KGaA MK_PG5156EM datasheet, 4 pages.
[cited by applicant]
Arnold, T.E. (Nov. 2005). “Fluid Purification Using Charge-Modified Depth Filtration Media,” BioProcess International pp. 44-49.
[cited by applicant]
European Office Action, dated May 17, 2023, for European Patent Application No. 21701079.2, 6 pages.
[cited by applicant]
Follman, D.K. et al. (2004). “Factorial Screening of Antibody Purification Processes Using Three Chromatography Steps Without Protein A,” J. Chromatogr. A 1024(1-2):79-85.
[cited by applicant]
GE Healthcare (Nov. 2013). “Instructions 71-7129-00 AF: Phenyl Sepharose High Performance; Butyl Sepharose High Performance,” 16 pages.
[cited by applicant]
Ghose, S. et al. (2013). “Purification of Monoclonal Antibodies by Hydrophobic Interaction Chromatography Under No-Salt Conditions,” Mabs. 5(5):795-800.
[cited by applicant]
Kishore, R.S.K. et al. (Feb. 2011). “Degradation of Polysorbates 20 and 80: Studies on Thermal Autoxidation and Hydrolysis,” J. Pharm. Sci. 100(2):721-731.
[cited by applicant]
Li, Y. (2017, e-pub. Apr. 13, 2017). “Effective Strategies for Host Cell Protein Clearance in Downstream Processing of Monoclonal Antibodies and Fc-Fusion Proteins,” Protein Expression and Purification 134:96-103.
[cited by applicant]
Lu, C. et al. (Jan./Feb. 2013). “Characterization of Monoclonal Antibody Size Variants Containing Extra Light Chains,” mAbs 5(1):102-113.
[cited by applicant]
Pall Life Sciences Data Sheet (Sep. 2004). “Pall Mustang S Capsules,” PELEH/02.SH/CS/09.2004 Datasheet, 4 pages.
[cited by applicant]
Tianjin, J. et al. (2018). “Research Progress in Purification Technology of Antibody Drugs in Process of Large-Scale Production,” China Academic Journal 35(10):6-11. English Abstract, 6 pages.
[cited by applicant]
Wang, S. et al. (2018, e-pub. Mar. 16, 2018). “Characterization of Product-Related Low Molecular Weight Impurities in Therapeutic Monoclonal Antibodies Using Hydrophilic Interaction Chromatography Coupled with Mass Spec…
[cited by applicant]
Xu, J. (2011). “Viral and Plasmid Transduction Systems: Methods to Modify Immune Cells for Cancer Immunotherapy,” Nature Biotechnology, 29 pages.
[cited by applicant]
Hester, J. et al. (Oct. 2020). “Streamlined Polishing and Viral Clearance: Using a New Hybrid, Biomimetic, Single-Use Anion Exchanger,” BioProcess International 18(10):70-76.
[cited by applicant]
MILLISTAK+ ® HC Pro (Sep. 9, 2020). Millipore, With English Translation. 12 pages.
[cited by applicant]