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Continuous Countercurrent Chromatography in Protein Purification

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Protein Chromatography

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2699))

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Abstract

Continuous countercurrent chromatography can be applied for both capture and polishing steps in the downstream processing of biopharmaceuticals. This chapter explains the concept of countercurrent operation, focusing on twin-column processes and how it can be used to alleviate the trade-offs of traditional batch chromatography with respect to resin utilization/productivity and yield/purity. CaptureSMB and MCSGP, the main twin-column continuous countercurrent chromatography processes, are explained, and the metrics by which they are compared to single-column chromatography are identified. Practical hints for process design and application examples are provided. Finally, regulatory aspects, scale-up, and UV-based process control are covered.

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References

  1. Konstantinov K (2013) The promise of continuous bioprocessing. Castelldefells, Spain, 20–24 Oct 2013

    Google Scholar 

  2. Mahajan E, George A, Wolk B (2012) Improving affinity chromatography resin efficiency using semi-continuous chromatography. J Chromatogr A 1227:154–162

    Article  CAS  PubMed  Google Scholar 

  3. Carta G, Jungbauer A (2010) Effects of dispersion and adsorption kinetics on column performance. In: Protein chromatography. Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, pp 237–276

    Chapter  Google Scholar 

  4. Godawat R et al (2012) Periodic counter-current chromatography – design and operational considerations for integrated and continuous purification of proteins. Biotechnol J 7(12):1496–1508

    Article  CAS  PubMed  Google Scholar 

  5. Warikoo V et al (2012) Integrated continuous production of recombinant therapeutic proteins. Biotechnol Bioeng 109(12):3018–3029

    Article  CAS  PubMed  Google Scholar 

  6. Baur D (2016) Comparison of batch and continuous multi-column protein A capture processes. J Biotechnol 11(7):920–931

    Article  CAS  Google Scholar 

  7. Angarita M et al (2015) Twin-column CaptureSMB: a novel cyclic process for protein A affinity chromatography. J Chromatogr A 1389:85–95

    Article  CAS  PubMed  Google Scholar 

  8. Ulmer N, Muller-Spath T, Neunstoecklin B, Aumann L, Bavand M, Morbidelli M (2015) Affinity capture of F(ab’)2 fragments: using twin-column countercurrent chromatography. Bioprocess Int 13(2):22–29

    Google Scholar 

  9. Pollock J et al (2013) Optimising the design and operation of semi-continuous affinity chromatography for clinical and commercial manufacture. J Chromatogr A 1284:17–27

    Article  CAS  PubMed  Google Scholar 

  10. Müller-Späth T et al (2008) Chromatographic separation of three monoclonal antibody variants using multicolumn countercurrent solvent gradient purification (MCSGP). Biotechnol Bioeng 100(6):1166–1177

    Article  PubMed  Google Scholar 

  11. Müller-Späth T et al (2010) Increasing the activity of monoclonal antibody therapeutics by continuous chromatography (MCSGP). Biotechnol Bioeng 107(4):652–662

    Article  PubMed  Google Scholar 

  12. Müller-Späth T, Morbidelli M (2014) Multicolumn countercurrent gradient chromatography for the purification of biopharmaceuticals. In: Continuous processing in pharmaceutical manufacturing. Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, pp 227–254

    Chapter  Google Scholar 

  13. Müller-Späth T, Ulmer N, Aumann L, Ströhlein G, Bavand M, Hendriks LJA, de Kruif J, Throsby M, Bakker ABH (2013) Purifying common light-chain bispecific antibodies: a twin-column, countercurrent chromatography platform process. Bioprocess Int 11(5):36–45

    Google Scholar 

  14. Müller-Späth T, Morbidelli M (2017) Countercurrent chromatography for the purification of monoclonal antibodies, bispecific antibodies, and antibody–drug conjugates. In: Process scale purification of antibodies. Wiley, Hoboken, pp 391–407

    Chapter  Google Scholar 

  15. Takizawa BT (2011) Evaluation of the financial impact of continuous chromatography in the production of biologics. http://hdl.handle.net/1721.1/66045

  16. Angelo J, Pagano J, Muller-Spath T, Mihlbachler K, Chollangi S, Xu X, Ghose S, Li ZJ (2018) Scale-up of twin-column periodic counter-current chromatography for MAb purification. Bioprocess Int 16(4):1–6

    Google Scholar 

  17. Janet W (2014) Modernizing pharmaceutical manufacturing – continuous manufacturing as a key enabler. In: International symposium on continuous manufacturing of pharmaceuticals, Cambridge

    Google Scholar 

  18. Baur D et al (2019) Model-assisted process characterization and validation for a continuous two-column protein A capture process. Biotechnol Bioeng 116(1):87–98

    Article  CAS  PubMed  Google Scholar 

  19. Angelo J et al (2019) Virus clearance validation across continuous capture chromatography. Biotechnol Bioeng 116(9):2275–2284

    Article  CAS  PubMed  Google Scholar 

  20. Angelo JM et al (2021) Continued insights into virus clearance validation across continuous capture chromatography. Biotechnol Bioeng 118(9):3604–3609

    Article  CAS  PubMed  Google Scholar 

  21. Brestrich N et al (2015) Advances in inline quantification of co-eluting proteins in chromatography: process-data-based model calibration and application towards real-life separation issues. Biotechnol Bioeng 112(7):1406–1416

    Article  CAS  PubMed  Google Scholar 

  22. Krättli M, Steinebach F, Morbidelli M (2013) Online control of the twin-column countercurrent solvent gradient process for biochromatography. J Chromatogr A 1293:51–59

    Article  PubMed  Google Scholar 

  23. Bigelow E et al (1643) Using continuous chromatography methodology to achieve high-productivity and high-purity enrichment of charge variants for analytical characterization. J Chromatogr A 2021:462008

    Google Scholar 

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Correspondence to Thomas Müller-Späth .

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Müller-Späth, T. (2023). Continuous Countercurrent Chromatography in Protein Purification. In: Loughran, S.T., Milne, J.J. (eds) Protein Chromatography. Methods in Molecular Biology, vol 2699. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3362-5_3

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  • DOI: https://doi.org/10.1007/978-1-0716-3362-5_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3361-8

  • Online ISBN: 978-1-0716-3362-5

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