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17–19 Sept 2018
Fakultät für Maschinenwesen der Technischen Universität München
Europe/Berlin timezone

Magnetic protein separation with new affinity tags for bare iron oxide nanoparticles

17 Sept 2018, 16:30
1h 30m
Fakultät für Maschinenwesen der Technischen Universität München

Fakultät für Maschinenwesen der Technischen Universität München

Boltzmannstraße 15 85748 Garching b. München
Poster P6 Nanomaterials and nanostructures Poster session 1

Speaker

Dr Sebastian Schwaminger (Bioseparation Engineering Group)

Description

The need for pharmaceuticals based on therapeutic proteins is increasing for many years. Here, the purification is the most cost-intensive step during the production of therapeutic proteins which leads to remarkable potentials in the improvement of common techniques. Besides adaption of purification resin, affinity tags can be used to selectively separate target proteins from contaminations. Commonly, such affinity peptide sequences only bind selectively to expensive surface modifications. Therefore, we design highly affine peptides, utilized as tags for magnetite nanoparticles.
Magnetite nanoparticles are synthesized with a co-precipitation route and used for protein purification without further modifications.
A glutamate tag is used as affinity sequence in a green fluorescent protein (GFP). High loads of GFP on the magnetite nanoparticles can be reached at different buffer conditions which can be verified by fluorescence analysis. However, the binding of the glutamate tag to the nanoparticles is reversible.
The colloidal behavior of the nanoparticles and their interaction with the proteins is investigated with small angle neutron scattering. Here, we are able to observe small aggregates of around 4 primary particles which tend to agglomerate when exposed to proteins and E. coli fermentation broths. However, this agglomeration does not affect the binding capacity while aiding the magnetic separation of nanoparticles. Thus, the developed particle-tag system can be up-scaled.

Primary authors

Dr Sebastian Schwaminger (Bioseparation Engineering Group) Dr Silvia Blank-Shim (Bioseparation Engineering Group) Dr Paula Fraga-García (Bioseparation Engineering Group) Ms Isabell Scheifele (Bioseparation Engineering Group) Dr Vitaliy Pipich (Jülich Centre for Neutron Science) Prof. Sonja Berensmeier (Bioseparation Engineering Group)

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