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End-to-end continuous production of complex recombinant proteins

End-to-end continuous production of complex recombinant proteins . Integration of perfusion cultivation and automated multi-step purification. Tiainen P., Bjelke J. R., Skibstrup D., Heitmann M. and Åkesson M. . Intro. A continuous and multi-step ÄKTA was developed

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End-to-end continuous production of complex recombinant proteins

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  1. Integration of perfusion cultivation and automated multi-step purification End-to-end continuous production of complex recombinant proteins Integration of perfusion cultivation and automated multi-step purification Tiainen P., Bjelke J. R., Skibstrup D., Heitmann M. and Åkesson M.

  2. Integration of perfusion cultivation and automated multi-step purification Intro A continuous and multi-step ÄKTA was developed No customised hardware or software An ATF perfusion cultivation system and the modified ÄKTA was integrated It will increase our capacity to supply research proteins And enable us to investigate continuous production approaches in a thorough way

  3. Integration of perfusion cultivation and automated multi-step purification The whole setup – ATF reactor coupled to an modified ÄKTA ATF perfusion system Modified ÄKTApure

  4. Integration of perfusion cultivation and automated multi-step purification Benefits of integrated production • Small foot print • Low residence time from protein expressed to protein in final formulation buffer • Low resin usage • Just-enough production • Low manual attendance need • Continuous monitoring of the cultivation performance

  5. Integration of perfusion cultivation and automated multi-step purification A close-up of the system

  6. Integration of perfusion cultivation and automated multi-step purification A close-up of the system Steady state, control of viable biomass

  7. Integration of perfusion cultivation and automated multi-step purification A close-up of the system

  8. Integration of perfusion cultivation and automated multi-step purification A close-up of the system Continuous application

  9. Integration of perfusion cultivation and automated multi-step purification A close-up of the system Multi-step purification

  10. Integration of perfusion cultivation and automated multi-step purification The first steps towards end-to-end processing – multi-step processing A system enabling a multi-column setup was created A sample was circulated from an AIEX column to an buffer-exchange column and back, showing PoC Modified ÄKTAexplorer

  11. Integration of perfusion cultivation and automated multi-step purification The first steps towards end-to-end processing – continuous application A system enabling continuous application was created Batch 2 Batch 4 Batch 6 Batch 8 Consistent (good) protein quality was seen in eluted batches Modified ÄKTAexplorer

  12. Integration of perfusion cultivation and automated multi-step purification A ÄKTApure can accomplish both continuous and multi-step processing Feature from a continuous Explorer Feature from a multi-step Explorer A continuous and a multi-step Pure

  13. Integration of perfusion cultivation and automated multi-step purification A continuous campaign A six-step process was made to run automated All on one system During the time one of the capture columns is loaded the other protein batch is fully purified Every column can be targeted with e.g. step-specific buffers and e.g. gradient elution

  14. Integration of perfusion cultivation and automated multi-step purification A continuous campaign A complete chromatogram is generated Purified protein

  15. Integration of perfusion cultivation and automated multi-step purification A screen dump of a on-going campaign 10 000 min

  16. Integration of perfusion cultivation and automated multi-step purification A continuous campaign All elution profiles can be found in the full chromatogram Note: Gradient elution

  17. Integration of perfusion cultivation and automated multi-step purification A continuous campaign – the modified ÄKTApure Note: Small columns

  18. Integration of perfusion cultivation and automated multi-step purification A continuous campaign – ATF reactor coupled to the modified ÄKTApure Continuous and multi-step Pure 5L ATF perfusion reactor

  19. Integration of perfusion cultivation and automated multi-step purification Closing remarks • A integrated and automated protein production process was established • No customised hardware or software was needed • It performs well

  20. Integration of perfusion cultivation and automated multi-step purification Thank you for your attention! Questions? Comments?

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