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A Different Perspective: How Much Innovation Is Really Needed for Monoclonal Antibody Production Using Mammalian Cell Technology?

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New Bioprocessing Strategies: Development and Manufacturing of Recombinant Antibodies and Proteins

Part of the book series: Advances in Biochemical Engineering/Biotechnology ((ABE,volume 165))

Abstract

As biopharmaceutical companies have optimized cell line and production culture process development, titers of recombinant antibodies have risen steadily to 3–8 g/L for fed-batch mammalian cultures at production scales of 10 kL or larger. Most new antibody products are produced from Chinese Hamster Ovary (CHO) cell lines, and there are relatively few alternative production hosts under active evaluation. Many companies have adopted a strategy of using the same production cell line for early clinical phases as well as commercial production, which reduces the risk of product comparability issues during the development lifecycle. Product quality and consistency expectations rest on the platform knowledge of the CHO host cell line and processes used for the production of many licensed antibodies. The lack of impact of low-level product variants common to this platform on product safety and efficacy also builds on the established commercial history of recombinant antibodies, which dates back to 1997.

Efforts to increase titers further will likely yield diminishing returns. Very few products would benefit significantly from a titer greater than 8 g/L; in many cases, a downstream processing bottleneck would preclude full recovery from production-scale bioreactors for high titer processes. The benefits of a process platform based on standard fed-batch production culture include predictable scale-up, process transfer, and production within a company’s manufacturing network or at a contract manufacturing organization. Furthermore, the confidence in an established platform provides key support towards regulatory flexibility (e.g., design space) for license applications following a quality-by-design strategy.

These factors suggest that novel technologies for antibody production may not provide a substantial return on investment. What, then, should be the focus of future process development efforts for companies that choose to launch antibody products using their current platform? This review proposes key focus areas in an effort to continually improve process consistency, assure acceptable product quality, and establish appropriate process parameter limits to enable flexible manufacturing options.

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References

  1. Nagy A (2000) Cre recombinase: the universal reagent for genome tailoring. Genesis 26:99–109

    CAS  PubMed  Google Scholar 

  2. Zhang L, Inniss M, Han S, Moffat M, Jones H, Zhang B, Cox W, Rance J, Young R (2015) Recombinase-mediated cassette exchange (RMCE) for monoclonal antibody expression in the commercially relevant CHOK1SV cell line. Biotechnol Prog 31(6):1645–1656

    CAS  PubMed  Google Scholar 

  3. Evans K, Albanetti T, Venkat R, Schoner R, Savery J, Miro-Quesada G, Rahan B, Groves C (2015) Assurance of monoclonality in one round of cloning through cell sorting for single cell deposition coupled with high resolution cell imaging. Biotechnol Prog 31(5):1172–1179

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Frye C, Deshpande R, Estes S, Francissen K, Joly J, Lubiniecki A, Munro T, Russell R, Wang T, Anderson K (2016) Industry view on the relative importance of “clonality” of biopharmaceutical-producing cell lines. Biologicals 44(2):117–122

    PubMed  Google Scholar 

  5. Chen A, Chitta R, Chang D, Amanullah A (2009) Twenty-four well plate miniature bioreactor systems as a scale-down model for cell culture process development. Biotechnol Bioeng 102(1):148–160

    CAS  PubMed  Google Scholar 

  6. Rouiller Y, Perilleux A, Collet N, Jordan M, Stettler M, Broly H (2013) A high-throughput media design approach for high performance mammalian fed-batch cultures. MAbs 5(3):501–511

    PubMed  PubMed Central  Google Scholar 

  7. Goochee C (2002) The role of a process development group in biopharmaceutical process startup. Cytotechnology 38:63–76

    PubMed  PubMed Central  Google Scholar 

  8. Pohlscheidt M, Corrales M, Charaniya S, Fallon E, Bruch M, Jenzsch M, Sieblist C (2013) Avoiding pitfalls during technology transfer of cell culture manufacturing processes in the pharmaceutical industry—mitigating risk and optimizing performance. Pharm Outsourcing 14:34–48

    Google Scholar 

  9. Hammond M, Marghitoiu L, Lee H, Perez L, Rogers G, Nashed-Samuel Y, Nunn H, Kline S (2014) A cytotoxic leachable compound from single-use bioprocess equipment that causes poor cell growth performance. Biotechnol Prog 30(2):332–337

    CAS  PubMed  Google Scholar 

  10. Horvath B, Tsang V, Lin W, Dai X-P, Kunas K, Frank G (2013) A generic growth test method for improving quality control of disposables in industrial cell culture. BioPharm Int 12(6):34–41

    Google Scholar 

  11. Wood J, Mahajan E, Shiratori M (2013) Strategy for selecting disposable bags for cell culture media applications based on a root-cause investigation. Biotechnol Prog 29(6):1535–1549

    CAS  PubMed  Google Scholar 

  12. Garnick R (1996) Experience with viral contamination in cell culture. Dev Biol Stand 88:49–56

    CAS  PubMed  Google Scholar 

  13. Moody M, Alves W, Varghese J, Khan F (2011) Mouse minute virus (MMV) contamination—a case study: detection, root cause determination, and corrective actions. PDA J Pharm Sci Technol 65(6):580–588

    PubMed  Google Scholar 

  14. Skrine J (2011) A biotech production facility contamination case study—minute mouse virus. PDA J Pharm Sci Technol 65(6):599–611

    PubMed  Google Scholar 

  15. Kiss R (2011) Practicing safe cell culture: applied process designs for minimizing virus contamination risk. PDA J Pharm Sci Technol 65(6):715–729

    PubMed  Google Scholar 

  16. Shiratori M, Kiss R (2017) Risk mitigation in preventing adventitious agent contamination of mammalian cell cultures. Adv Biochem Eng Biotechnol. https://doi.org/10.1007/10_2017_38

  17. Gramer M (2013) Product quality considerations for mammalian cell culture process development and manufacturing. In: Zhou W, Kantardjieff A (eds) Mammalian cell cultures for biologics manufacturing. Advances in biochemical engineering/biotechnology, vol 139. Springer, Berlin

    Google Scholar 

  18. Hossler P, McDermott S, Racicot C, Fann J (2013) Improvement of mammalian cell culture performance through surfactant enabled concentrated feed media. Biotechnol Prog 29(4):1023–1033

    CAS  PubMed  Google Scholar 

  19. Luo J, Zhang J, Ren D, Tsai WL, Li F, Amanullah A, Hudson T (2012) Probing of C-terminal lysine variation in a recombinant monoclonal antibody production using Chinese hamster ovary cells with chemically defined media. Biotechnol Bioeng 109(9):2306–2315

    CAS  PubMed  Google Scholar 

  20. Vijayasankaran N, Varma S, Yang Y, Mun M, Arevalo S, Gawlitzek M, Swartz T, Lim A, Li F, Zhang B, Meier S, Kiss R (2013) Effect of cell culture medium components on color of formulated monoclonal antibody drug substance. Biotechnol Prog 29(5):1270–1277

    CAS  PubMed  Google Scholar 

  21. Yuk I (2014) Effects of copper on CHO cells: insights from gene expression analyses. Biotechnol Prog 30(2):429–442

    CAS  PubMed  Google Scholar 

  22. Kao Y-H, Hewitt D, Trexler-Schmidt M, Laird M (2010) Mechanism of antibody reduction in cell culture production processes. Biotechnol Bioeng 107(4):622–632

    CAS  PubMed  Google Scholar 

  23. Mun M, Khoo S, Do Minh A, Dvornicky J, Trexler-Schmidt M, Kao Y-H, Laird M (2015) Air sparging for prevention of antibody disulfide bond reduction in harvested CHO cell culture fluid. Biotechnol Bioeng 112(4):734–742

    CAS  PubMed  Google Scholar 

  24. Trexler-Schmidt M, Sargis S, Chiu J, Sze-Khoo S, Mun M, Kao Y-H, Laird M (2010) Identification and prevention of antibody disulfide bond reduction during cell culture manufacturing. Biotechnol Bioeng 106(3):452–461

    CAS  PubMed  Google Scholar 

  25. Kelley B (2009) Industrialization of mAb production technology: the bioprocessing industry at a crossroads. MAbs 1(5):443–452

    PubMed  PubMed Central  Google Scholar 

  26. Hossler P, Khattak S, Jian Z (2009) Optimal and consistent protein glycosylation in mammalian cell culture. Glycobiology 19(9):936–949

    CAS  PubMed  Google Scholar 

  27. Peng H, Ali A, Lanan M, Hughes E, Wiltberger K, Guan B, Prajapati S, Hu W (2016) Mechanism investigation for poloxamer 188 raw material variation in cell culture. Biotechnol Prog 32(3):767–775

    CAS  PubMed  Google Scholar 

  28. Gilbert A, Huang Y-M, Ryll T (2014) Identifying and eliminating cell culture process variability. Pharm Bioprocess 2(6):519–534

    Google Scholar 

  29. Kolwyck D (2013) How defined is chemically defined medium? IBC’s biopharmaceutical development and production week, Huntington Beach, CA, February 2013

    Google Scholar 

  30. Toro A, Colon J, Melendez-Colon V, Rivera J (2010) Changes in raw material sources from suppliers: determining their impact on customers’ biopharmaceutical manufacturing operations. Bioprocess Int 2010:50–55

    Google Scholar 

  31. Kirdar A, Green K, Rathore A (2008) Application of multivariate data analysis for identification and successful resolution of a root cause for a bioprocessing application. Biotechnol Prog 24(3):720–726

    CAS  PubMed  Google Scholar 

  32. Shi H, Goudar C (2014) Recent advances in the understanding of biological implications and modulation methodologies of monoclonal antibody N-linked high mannose glycans. Biotechnol Bioeng 111(10):1907–1919

    CAS  PubMed  Google Scholar 

  33. Zupke C, Brady L, Slade P, Clark P, Caspary R, Livinston B, Taylor L, Bigham K, Morris A, Bailey R (2015) Real-time product attribute control to manufacture antibodies with defined N-linked glycan levels. Biotechnol Prog 31:1433–1441

    CAS  PubMed  Google Scholar 

  34. Berry B, Moretto J, Matthews J, Smelko J, Wiltberger K (2015) Cross-scale predictive modeling of CHO cell culture growth and metabolites using Raman spectroscopy and multivariate analysis. Biotechnol Prog 31(2):566–577

    CAS  PubMed  Google Scholar 

  35. Matthews T, Berry B, Smelko J, Moretto J, Moore B, Wiltberger K (2016) Closed loop control of lactate concentration in mammalian cell culture by Raman spectroscopy leads to improved cell density, viability, and biopharmaceutical protein production. Biotechnol Bioeng 113(11):2416–2124

    CAS  PubMed  Google Scholar 

  36. Tai M, Ly A, Leung I, Nayar G (2015) Efficient high-throughput biological process characterization: definitive screening design with the ambr250 bioreactor system. Biotechnol Prog 31(5):1388–1395

    CAS  PubMed  Google Scholar 

  37. ICH International Conference on Harmonization Q8, Pharmaceutical Development (2009)

    Google Scholar 

  38. Luciani F, Galluzzo S, Gaggioli A, Kruse N, Venneugues P, Schneider C, Pini C, Melchiorri D (2015) Implementing quality by design for biotech products: are regulators on track? MAbs 7(3):451–455

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Hakemeyer C, McKnight N, St. John R, Meier S, Trexler-Schmidt M, Kelley B, Zettl F, Puskeiler R, Kleinjans A, Lim F, Wurth C (2016) Process characterization and design space definition. Biologicals 44(5):306–318

    PubMed  Google Scholar 

  40. Janakiraman V, Kwiatkowski C, Kshirsagar R, Ryll T, Huang Y-M (2015) Application of high-throughput mini-bioreactor system for systematic scale-down modeling, process characterization, and control strategy development. Biotechnol Prog 31(6):1623–1632

    CAS  PubMed  Google Scholar 

  41. Rameez S, Mostafa S, Miller C, Shukla A (2014) High-throughput miniaturized bioreactors for cell culture process development: reproducibility, scalability, and control. Biotechnol Prog 30(3):718–727

    CAS  PubMed  Google Scholar 

  42. Goudar C, Chen C, Le H (2015) Biopharmaceuticals—continuous processing in upstream operations. Chemical Engineering Progress, New York

    Google Scholar 

  43. Croughan MS, Konstantinov KB, Cooney C (2015) The future of industrial bioprocessing: batch or continuous? Biotechnol Bioeng 112(4):648–651

    CAS  PubMed  Google Scholar 

  44. Hernandez R (2015) Continuous manufacturing: a changing processing paradigm. BioPharm Int 28(4):20–27

    Google Scholar 

  45. Farid SS, Pollock J, Ho SV (2014) Evaluating the economic and operational feasibility of continuous processes for monoclonal antibodies. Continuous processing in pharmaceutical manufacturing. Wiley, Weinheim, pp 433–456

    Google Scholar 

  46. Zydney A (2016) Continuous downstream processing for high value biological products: a review. Biotechnol Bioeng 113(3):465–475

    CAS  PubMed  Google Scholar 

  47. Jagschies G (2012) Changing upstream development to improve the downstream process and the overall yield and product quality, IBC life sciences conference, monoclonal antibody development & production, San Diego

    Google Scholar 

  48. Simmons L, Reilly D, Klimowski L, Raju T, Meng G, Sims P, Hong K, Shields R, Damico L, Rancatore P, Yansura D (2002) Expression of full-length immunoglobulins in Escherischia coli: rapid and efficient production of aglycosylated antibodies. J Immunol Methods 263(1–2):133–147

    CAS  PubMed  Google Scholar 

  49. Grampp G, Ramanan S (2013) Managing unexpected events in the manufacturing of biologic medicines. BioDrugs 27(4):305–316

    PubMed  Google Scholar 

  50. Turner R, Joseph A, Titchener-Hooker N, Bender J (2016) Manufacturing of proteins and antibodies: chapter downstream processing technologies—harvest operations. Adv Biochem Eng Biotechnol. https://doi.org/10.1007/10_2016_54

  51. Smelko J, Wiltberger K, Hickman E, Morris B, Blackburn T, Ryll T (2011) Performance of high intensity fed-batch mammalian cell cultures in disposable bioreactor systems. Biotechnol Prog 27(5):1358–1364

    CAS  PubMed  Google Scholar 

  52. Yang W, Minklera D, Kshirsagar R, Ryll T, Huang Y-M (2016) Concentrated fed-batch cell culture increases manufacturing capacity without additional volumetric capacity. J Biotechnol 217:1–11

    CAS  PubMed  Google Scholar 

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Acknowledgements

Numerous technical staff at Genentech and Roche have contributed over the years to the collective platform process and the understanding of its capabilities, its historical challenges, and the opportunities for future improvements briefly summarized in this chapter. In addition, further acknowledgement is warranted to the many individuals and companies within the biotechnology industry that have driven the advances responsible for making the mammalian cell culture processes for antibody production the highly productive systems they are today.

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Authors and Affiliations

  1. Vir Biotechnology, Inc., San Francisco, CA, USA

    Brian Kelley

  2. Sutro Biopharma, Inc., San Francisco, CA, USA

    Robert Kiss

  3. Genentech (A Member of the Roche Group), San Francisco, CA, USA

    Michael Laird

Authors
  1. Brian Kelley
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  2. Robert Kiss
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  3. Michael Laird
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Corresponding author

Correspondence to Brian Kelley .

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Editors and Affiliations

  1. Sutro Biopharma Inc., San Francisco, CA, USA

    Bob Kiss

  2. Lonza, Basel, Switzerland

    Uwe Gottschalk

  3. Solothurn Manufacturing Facility, Biogen International GmbH, Luterbach, Switzerland

    Michael Pohlscheidt

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Kelley, B., Kiss, R., Laird, M. (2018). A Different Perspective: How Much Innovation Is Really Needed for Monoclonal Antibody Production Using Mammalian Cell Technology?. In: Kiss, B., Gottschalk, U., Pohlscheidt, M. (eds) New Bioprocessing Strategies: Development and Manufacturing of Recombinant Antibodies and Proteins. Advances in Biochemical Engineering/Biotechnology, vol 165. Springer, Cham. https://doi.org/10.1007/10_2018_59

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