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End-to-end continuous production of complex recombinant proteins
Integration of perfusion cultivation and automated multi-step purification
Integration of perfusion cultivation and automated multi-step purification 2013-10-22 1
Tiainen P., Bjelke J. R., Skibstrup D., Heitmann M. and Åkesson M.
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 approachesin a thorough way
2013-10-22
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
2013-10-22
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
2013-10-22
Integration of perfusion cultivation and automated multi-step purification 5
A close-up of the system
2013-10-22
Integration of perfusion cultivation and automated multi-step purification 6
A close-up of the system
2013-10-22
Steady state, control of viable biomass
Integration of perfusion cultivation and automated multi-step purification 7
A close-up of the system
2013-10-22
Integration of perfusion cultivation and automated multi-step purification 8
Continuous application
A close-up of the system
2013-10-22
Integration of perfusion cultivation and automated multi-step purification 9
Multi-step purification
A close-up of the system
2013-10-22
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
2013-10-22
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 6 Batch 8
2013-10-22
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
2013-10-22
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
2013-10-22
Integration of perfusion cultivation and automated multi-step purification 14
A complete chromatogram is generated
Purified protein
A continuous campaign
2013-10-22
Integration of perfusion cultivation and automated multi-step purification 15
10 000 min
A screen dump of a on-going campaign
2013-10-22
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
2013-10-22
Integration of perfusion cultivation and automated multi-step purification 17
Note: Small columns
A continuous campaign – the modified ÄKTApure
2013-10-22
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
2013-10-22
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
2013-10-22
Integration of perfusion cultivation and automated multi-step purification 20
Thank you for your attention!
Questions? Comments?
2013-10-22