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High-Throughput Process Development for the Chromatographic Purification of Viral Antigens

Part of the Methods in Molecular Biology book series (MIMB,volume 2183)

Abstract

Chromatography is a widely used method in the biotechnology industry and functions to separate the desired product from process and product related impurities. There is a multitude of resins available based on different modalities (such as charge, hydrophobicity, and affinity) to provide a spectrum of approaches to meet the separation challenges of the diverse products. The challenge of developing viral antigen purification processes is addressed in this method. A unique feature of this product class is that in order to protect against more than one strain of an antigen, vaccines are often multivalent. This entails multiple production processes for each antigen, all of which will require separate development and validation. Ideally, a universal purification method is sought, but differences in the protein subunits (frequently used as the antigens) make this challenging and often-bespoke purification steps are required. This means process development for the chromatographic stages of these products can be particularly challenging and labour intensive. With the numerous choices available, making critical process decisions that are usually unique to each product, process, and strain, can be costly and time-consuming. To address this, scale down purification at <1.0 mL column volume and automation approaches are increasingly applied to increase throughput. In this work, a method is described wherein a Tecan Freedom EVO® automated liquid handler is deployed for the evaluation of different resin chemistries and buffer conditions to find a suitable purification strategy. This method allows for the rapid evaluation of the separation viral antigens where limited information on chromatography behavior is known at the early stages of process development. Here, we demonstrate the methodology firstly by explaining the automated purification script and secondly by applying the script for an efficient purification development for different serotypes of rotavirus antigens.

Key words

  • Process development
  • High-throughput process development (HTPD)
  • Rotavirus antigens
  • Purification
  • Chromatography

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References

  1. Bhambure R, Rathore A (2013) Chromatography process development in the quality by design paradigm I: establishing a high-throughput process development platform as a tool for estimating “characterization space” for an ion exchange chromatography step. Biotechnol Prog 29:403–414

    CAS  CrossRef  Google Scholar 

  2. Daniels C, Rodriguez J, Lim E et al (2016) An integrated robotic system for high-throughput process development of cell and virus culture conditions: application to biosafety level 2 live virus vaccines. Eng Life Sci 16:202–209

    CAS  CrossRef  Google Scholar 

  3. Wenger MD, Dephillips P, Price CE et al (2007) An automated microscale chromatographic purification of virus-like particles as a strategy for process development. Biotechnol Appl Biochem 47:131–139

    CAS  CrossRef  Google Scholar 

  4. Konstantinidis S, Welsh JP, Roush DJ et al (2016) Application of simplex-based experimental optimization to challenging bioprocess development problems: case studies in downstream processing. Biotechnol Prog 32:404–419

    CAS  CrossRef  Google Scholar 

  5. Konstantinidis S, Welsh JP, Titchener-Hooker NJ et al (2018) Data-driven multi-objective optimization via grid compatible simplex technique and desirability approach for challenging high throughput chromatography applications. Biotechnol Prog 34:1393–1406

    CAS  CrossRef  Google Scholar 

  6. Treier K, Hansen S, Richter C et al (2012) High-throughput methods for miniaturization and automation of monoclonal antibody purification processes. Biotechnol Prog 28:723–732

    CAS  CrossRef  Google Scholar 

  7. Welsh JP, Petroff MG, Rowicki P et al (2014) A practical strategy for using miniature chromatography columns in a standardized high-throughput workflow for purification development of monoclonal antibodies. Biotechnol Prog 30:626–635

    CAS  CrossRef  Google Scholar 

  8. Feliciano J, Berrill A, Ahnfelt M et al (2016) Evaluating high-throughput scale-down chromatography platforms for increased process understanding. Eng Life Sci 16:169–178

    CAS  CrossRef  Google Scholar 

  9. Łącki KM (2014) High throughput process development in biomanufacturing. Curr Opin Chem Eng 6:25–32

    CrossRef  Google Scholar 

  10. Konstantinidis S, Goh HY, Martin Bufájer JM et al (2018) Flexible and accessible automated operation of miniature chromatography columns on a liquid handling station. Biotechnol J 13:1700390

    CrossRef  Google Scholar 

  11. Li X, De Roo G, Burgers K et al (2012) Self-packed filter plates: a good alternative for pre-packed filter plates for developing purification processes for therapeutic proteins. Biotechnol J 7:1269–1276

    CrossRef  Google Scholar 

  12. Jacob SI, Khogeer B, Bampos N et al (2017) Development and application of synthetic affinity ligands for the purification of ferritin-based influenza antigens. Bioconjug Chem 28:1931–1943

    CAS  CrossRef  Google Scholar 

  13. El Khoury G, Wang Y, Wang D et al (2013) Design, synthesis, and assessment of a de novo affinity adsorbent for the purification of recombinant human erythropoietin. Biotechnol Bioeng 110:3063–3069

    CrossRef  Google Scholar 

  14. Khoury GE, Khogeer B, Chen C et al (2015) Bespoke affinity ligands for the purification of therapeutic proteins. Pharmaceut Bioprocess 3:139–152

    CAS  CrossRef  Google Scholar 

  15. Mcgown EL, Hafeman DG (1998) Multichannel pipettor performance verified by measuring pathlength of reagent dispensed into a microplate. Anal Biochem 258:155–157

    CAS  CrossRef  Google Scholar 

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Correspondence to Daniel G. Bracewell .

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Appendices

5. Appendix 1: EVOware Script for Preparation of Gradient Elution Buffers

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6. Appendix 2: EVOware Script for Performing RoboColumn Chromatography Using 0.6 mL Columns

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Jacob, S.I., Konstantinidis, S., Bracewell, D.G. (2021). High-Throughput Process Development for the Chromatographic Purification of Viral Antigens. In: Pfeifer, B.A., Hill, A. (eds) Vaccine Delivery Technology. Methods in Molecular Biology, vol 2183. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0795-4_9

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  • DOI: https://doi.org/10.1007/978-1-0716-0795-4_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0794-7

  • Online ISBN: 978-1-0716-0795-4

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