Transcript End-to-end continuous production of complex recombinant

Integration of perfusion cultivation and automated multi-step purification
2013-10-22
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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.
Integration of perfusion cultivation and automated multi-step purification
2013-10-22
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
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Integration of perfusion cultivation and automated multi-step purification
The whole setup – ATF reactor coupled to an modified ÄKTA
ATF perfusion system
Modified
ÄKTApure
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Integration of perfusion cultivation and automated multi-step purification
Benefits of integrated production
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Small foot print
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Low residence time from protein expressed
to protein in final formulation buffer
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Low resin usage
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Just-enough production
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Low manual attendance need
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Continuous monitoring of the cultivation
performance
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Integration of perfusion cultivation and automated multi-step purification
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A close-up of the system
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Integration of perfusion cultivation and automated multi-step purification
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A close-up of the system
Steady state, control of viable biomass
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Integration of perfusion cultivation and automated multi-step purification
2013-10-22
A close-up of the system
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Integration of perfusion cultivation and automated multi-step purification
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A close-up of the system
Continuous
application
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Integration of perfusion cultivation and automated multi-step purification
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A close-up of the system
Multi-step
purification
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Integration of perfusion cultivation and automated multi-step purification
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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
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Integration of perfusion cultivation and automated multi-step purification
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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
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Integration of perfusion cultivation and automated multi-step purification
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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
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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
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Integration of perfusion cultivation and automated multi-step purification
A continuous campaign
A complete chromatogram is generated
Purified
protein
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Integration of perfusion cultivation and automated multi-step purification
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A screen dump of a on-going campaign
10 000 min
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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
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Integration of perfusion cultivation and automated multi-step purification
A continuous campaign – the modified ÄKTApure
Note: Small columns
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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
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Integration of perfusion cultivation and automated multi-step purification
Closing remarks
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A integrated and automated protein
production process was established
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No customised hardware or software was
needed
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It performs well
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Integration of perfusion cultivation and automated multi-step purification
Thank you for your attention!
Questions? Comments?
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