Novel Strategy for Protein Purification

Recombinant proteins are used ubiquitously for medicinal, industrial and research purposes and purifying these proteins can be a costly, lengthy and complicated process. When pure protein is required, affinity chromatography is often used as the first step in a multi-step process, where a target protein is removed from cellular proteins through the affinity for a fixed medium. This requires either the target protein to have inherent affinity or the use of an affinity tag to introduce affinity. There are several commonly used tagging systems, each with their own strengths and weaknesses, although currently none can yield pure protein and must be followed with additional purification techniques such as ion exchange or gel filtration to clean up the protein. This aim of this PhD is to create a bacterial based tagging system using neuronal SNARE proteins. This group of proteins enable the release of neurotransmitters from neuronal cells by the formation of a protein complex which displays exceptionally high affinity and stability. The distinctive properties of this complex will be used to create a tagging system which will provide a one-step procedure to pure protein.