Skip to main content

Advertisement

SpringerLink
Log in

A Stable CHO K1 Cell Line for Producing Recombinant Monoclonal Antibody Against TNF-α

  • Original Paper
  • Published:
Molecular Biotechnology Aims and scope Submit manuscript

Abstract

Monoclonal antibodies (mAbs) are one of the most significant molecules in protein therapeutics. They are employed in the field of immunology, oncology and organ transplant. They have been also been employed for alleviating several bacterial and viral infections. Moreover, they have revolutionized the area of targeted therapy and improved the quality of treatments, as compared to other cytotoxic drugs and therapies. mAbs bind to specific molecules on the antigen and exhibit specificity towards that molecule, i.e. epitope. Thus, mAbs have immense opportunity to be explored for personalized therapy. The introduction of targeted mAb-based therapeutics has promoted many important scientific achievements in rheumatology. This has warranted additional investigations for developing newer mAb producing clones, to supplement the limited industrial production of certain mAb therapeutics. In this investigation, an integrative approach comprising optimized expression, selection and expansion was adopted to develop a mammalian cell line expressing mAb against TNF-α.The resulting stable clone is anticipated to serve as an economic alternative to the industrial clones, especially for research purposes. The clone was constructed for development of biosimilar of the highly valued therapeutic antibody, Humira.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

Abbreviations

mAb:

Monoclonal antibody

TNF- α:

Tumor necrosis factor alpha

CHO-K1:

Chinese hamster ovary –K1

G418:

Geneticin

FBS:

Fetal bovine serum

BSA:

Bovine serum albumin

ADCC:

Antibody dependent cell mediated cytoxicity

HRP:

Horseradish peroxidase

ELISA:

Enzyme-linked immunosorbent assay

HT:

Hypoxanthine

TMB:

Tetramethylenebenzidine

PCD:

Picogram protein/cell/day

HC:

Heavy chain

LC:

Light chain

IRES:

Internal ribosome entry site

FITC:

Fluorescein isothiocyanate

DAPI:

4′,6-Diamidino-2-phenylindole

IgG:

Immunoglobulin

Fc:

Fragment crystallizable region

References

  1. Gronemeyer, P., Ditz, R., & Strube, J. (2014). Trends in upstream and downstream process development for antibody manufacturing. Bioengineering, 1(4), 188–212

    Article  Google Scholar 

  2. Gupta, K., et al. (2019). Vector-related stratagems for enhanced monoclonal antibody production in mammalian cells. Biotechnology Advances

  3. Chames, P., et al. (2009). Therapeutic antibodies: Successes, limitations and hopes for the future. British Journal of Pharmacology, 157(2), 220–233

    Article  CAS  Google Scholar 

  4. Voronina, E., et al. (2016). Design of a stable cell line producing a recombinant monoclonal anti-TNFα antibody based on a CHO cell line. Springerplus, 5(1), 1584

    Article  CAS  Google Scholar 

  5. Alonso-Ruiz, A., et al. (2008). Tumor necrosis factor alpha drugs in rheumatoid arthritis: Systematic review and metaanalysis of efficacy and safety. BMC Musculoskeletal Disorders, 9(1), 52

    Article  Google Scholar 

  6. Kunert, R., & Reinhart, D. (2016). Advances in recombinant antibody manufacturing. Applied Microbiology and Biotechnology, 100(8), 3451–3461

    Article  CAS  Google Scholar 

  7. Azevedo, V. F., et al. (2016). Adalimumab: A review of the reference product and biosimilars. Biosimilars, 6, 29

    Article  CAS  Google Scholar 

  8. Furst, D., et al. (2007). (2007) Updated consensus statement on biological agents for the treatment of rheumatic diseases. Annals of the Rheumatic Diseases, 66(3), iii 2-iii22

    CAS  Google Scholar 

  9. Scott, D., & Kingsley, G. (2006). Tumor necrosis factor inhibitors for rheumatoid arthritis. New England Journal of Medicine, 355(7), 704–712

    Article  CAS  Google Scholar 

  10. Li, F., Shen, A., & Amanullah, A. (2010). Cell culture processes in monoclonal antibody production. (pp. 1–38). Pharmaceutical Sciences Encyclopedia: Drug Discovery, Development, and Manufacturing.

    Google Scholar 

  11. Costa, A. R., et al. (2010). Guidelines to cell engineering for monoclonal antibody production. European Journal of Pharmaceutics and Biopharmaceutics, 74(2), 127–138

    Article  Google Scholar 

  12. Xu, X., et al. (2011). The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line. Nature biotechnology, 29(8), 735

    Article  CAS  Google Scholar 

  13. Wurm, F. M. (2013). CHO quasispecies—Implications for manufacturing processes. Processes, 1(3), 296–311

    Article  Google Scholar 

  14. Sinici, I., et al. (2006). Comparison of HCMV IE and EF-1 promoters for the stable expression of β-subunit of hexosaminidase in CHO cell lines. Biochemical Genetics, 44(3–4), 168–175

    Article  Google Scholar 

  15. Kober, L., Zehe, C., & Bode, J. (2013). Optimized signal peptides for the development of high expressing CHO cell lines. Biotechnology and Bioengineering, 110(4), 1164–1173

    Article  CAS  Google Scholar 

  16. Sandbichler, A. M., Aschberger, T., & Pelster, B. (2013). A method to evaluate the efficiency of transfection reagents in an adherent zebrafish cell line. BioResearch Open Access, 2(1), 20–27

    Article  CAS  Google Scholar 

  17. Harlow, E., & Lane, D. (1999). Using Antibodies Cold Spring Harbor Lab Press. Cold Spring Harbor Press.

  18. Lipman, N. S., et al. (2005). Monoclonal versus polyclonal antibodies: Distinguishing characteristics, applications, and information resources. ILAR Journal, 46(3), 258–268

    Article  CAS  Google Scholar 

  19. Gross, A., et al. (2015). Technologies for single-cell isolation. International Journal of Molecular Sciences, 16(8), 16897–16919

    Article  CAS  Google Scholar 

  20. Ozturk, S. S., & Palsson, B. O. (1991). Physiological changes during the adaptation of hybridoma cells to low serum and serum-free media. Biotechnology and Bioengineering, 37(1), 35–46

    Article  CAS  Google Scholar 

  21. Rodrigues, M. E., et al. (2013). Advances and drawbacks of the adaptation to serum-free culture of CHO-K1 cells for monoclonal antibody production. Applied Biochemistry and Biotechnology, 169(4), 1279–1291

    Article  CAS  Google Scholar 

  22. Reinhart, D., et al. (2015). Benchmarking of commercially available CHO cell culture media for antibody production. Applied Microbiology and Biotechnology, 99(11), 4645–4657

    Article  CAS  Google Scholar 

  23. Pan, X., et al. (2017). Selection of chemically defined media for CHO cell fed-batch culture processes. Cytotechnology, 69(1), 39–56

    Article  CAS  Google Scholar 

  24. Lee, J. J., et al. (2019). Demonstration of functional similarity of a biosimilar adalimumab SB5 to Humira®. Biologicals, 58, 7–15

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Kritika Gupta is thankful to Indian Council of Medical Research(ICMR), No. 3/1/3/JRF-2015/HRD-LS/29/31232/85 for fellowship and financial support. Deepak Modi is thankful to ICMR for financial support. Prajakta Dandekar is thankful to Department of Science and Technology (DST), Science and Engineering Research Board (SERB): Ramanujan Fellowship Grant (SR/S2/RJN-139/2011) and UGC for Start-up Grant20-1/2012 (BSR)/20-1(10)/2012(BSR). Ratnesh Jain is thankful to DBT, Ministry of Science and Technology: Ramalingaswami Fellowship Grant (BT/RLF/Re-entry/51/2011).

Author information

Authors and Affiliations

  1. Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Mumbai, 400019, India

    Kritika Gupta & Prajakta Dandekar

  2. Department of Chemical Engineering, Institute of Chemical Technology, Mumbai, 400019, India

    Kritika Gupta & Ratnesh Jain

  3. Department of Molecular and Cellular Biology, ICMR-National Institute for Research in Reproductive Health, Mumbai, 400012, India

    Deepak Modi

Authors
  1. Kritika Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Deepak Modi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ratnesh Jain
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Prajakta Dandekar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Ratnesh Jain or Prajakta Dandekar.

Ethics declarations

Conflict of interest

All the contributing authors have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Gupta, K., Modi, D., Jain, R. et al. A Stable CHO K1 Cell Line for Producing Recombinant Monoclonal Antibody Against TNF-α. Mol Biotechnol 63, 828–839 (2021). https://doi.org/10.1007/s12033-021-00329-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12033-021-00329-4

Keywords

Search

Navigation