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

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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

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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

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Integration of perfusion cultivation and automated multi-step purification 5

A close-up of the system

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Integration of perfusion cultivation and automated multi-step purification 6

A close-up of the system

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Steady state, control of viable biomass

Integration of perfusion cultivation and automated multi-step purification 7

A close-up of the system

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Continuous application

A close-up of the system

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Multi-step purification

A close-up of the system

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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

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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

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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

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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

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A complete chromatogram is generated

Purified protein

A continuous campaign

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10 000 min

A screen dump of a on-going campaign

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All elution profiles can be found in the full chromatogram

Note: Gradient elution

A continuous campaign

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Note: Small columns

A continuous campaign – the modified ÄKTApure

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5L ATF perfusion reactor

Continuous and multi-step Pure

A continuous campaign – ATF reactor coupled to the modified ÄKTApure

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• A integrated and automated protein production process was established

• No customised hardware or software was needed

• It performs well

Closing remarks

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Thank you for your attention!Questions? Comments?

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