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End-to-end continuous production of complex recombinant proteins Integration of perfusion cultivation and automated multi-step purification 2013-10-22.

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Presentation on theme: "End-to-end continuous production of complex recombinant proteins Integration of perfusion cultivation and automated multi-step purification 2013-10-22."— Presentation transcript:

1 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 2 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 3 ATF perfusion system Modified ÄKTApure The whole setup – ATF reactor coupled to an modified ÄKTA

4 Integration of perfusion cultivation and automated multi-step purification 4 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 Benefits of integrated production

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

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

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

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

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

10 Integration of perfusion cultivation and automated multi-step purification 10 The first steps towards end-to-end processing – multi-step processing Modified ÄKTAexplorer 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

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

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

13 Integration of perfusion cultivation and automated multi-step purification 13 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 A continuous campaign

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

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

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

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

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

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

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


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