NERS · Novel Electro-Responsive Protein Separation Method with Magnetic Nanoparticles

Novel Electro-Responsive Protein Separation Method with Magnetic Nanoparticles

Purification is a critical step in drug manufacturing, which helps minimise undesired materials that compromise drug efficacy. Protein tags – peptide sequences attached to proteins – are commonly used to detect and purify expressed proteins. The EU-funded NERS project will explore more efficient protein purification approaches to help retain properties of pharmaceutically relevant proteins such as antibodies. It will extend prior work on the design of protein tags that had high binding affinity and selectivity for iron oxide magnetic materials. The project will develop an elution process that will help separate proteins containing peptide tags from those interacting with electric fields. Pharmaceuticals for cancer therapies and other diseases are very since the production of therapeutic molecules such as antibodies is costly and every production process needs to be developed individually. Especially purification processes, which make up to 80-90% of the whole production, need to be improved or new ideas need to be developed. Short peptide sequences, so called “tags”, can be used to create new purification strategies based on the biomolecule recognition of these sequences. Magnetic iron oxide nanoparticles are an interesting counterpart for peptide tags as their properties facilitate an easy handling and manipulation. I developed such a magnetite-binding peptide tag which allows the purification of tagged model proteins from fermentation broths by changing the surrounding media. However, such pH and buffer switches might also alter the properties of pharmaceutically relevant proteins such as antibodies. The challenge of this project is to establish a novel elution process based on an electrical potential switch instead of a pH switch. The process contains the magnetic separation of proteins containing the peptide tags and the elution of proteins based on the change of tag-particle interactions with electric fields. The use of this system will help to understand the binding of proteins to iron oxide nanoparticles and the formation of an electrochemical double layer in external fields. The electrical double layer formation is not only interesting in biotechnological processes but for the understanding of electrochemical catalysis and energy storage. This idea might pave the way to completely new approaches in biomolecule recognition, protein detection and purification. Objectives: 1. The development of a suitable electrode set-up to which magnetic nanoparticles can be magnetically transported and where a potential can be applied which influences the electrochemical double layer around the magnetic nanoparticles. This set-up represents the proof of principle for influencing the electrochemical double layer of nanomaterials magnetically deposited to an electrode and will be the basis for further improvements and applications. 2. The evaluation and development of suitable nanomaterials for the binding of tagged proteins, the magnetic separation and the control of the electrochemical double layer with a potential switch at the electrode set-up. This objective aims to prove the possibility to bind and elute proteins by a potential switch in a similar manner as by a pH shift. 3. The application and exploitation of this technique for new processes. Investigation of the protein separation and purification principles under “real” conditions in complex media such as crude cell lysates.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз