The production of biotherapeutics including antibodies and antibody fragments is a rapidly expanding market with an increasing number of products being approved for use. One of the major platforms used for production of such therapeutics is Escherichia coli, which offers a rapid production at low production costs. The favoured location for targeting these biotherapeutic is the periplasm of E. coli as this environment supports the formation of disulphide bonds and simplifies the purification process. There are a number of periplasmic release procedures currently practised in industry including osmotic shock. However their limitations call for the development of an improved generic periplasmic release method. This project demonstrates how the polymer poly(styrene-co-maleic acid) (SMA) can be applied as a novel and alternative periplasmic release agent. The amphipathic polymer self-assembles into discs encapsulating membrane proteins and thereby destabilises the outer membrane consequently releasing the periplasm. Data presented here show that SMA releases the model target proteins with a higher yield at equal or higher target purity than the conventional methods. Furthermore the developed methods was analysed and refined to be compatible with existing downstream and first steps for its adaptation on industrial scale were made.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:720714 |
| Date | January 2017 |
| Creators | Krämer, Julia |
| Publisher | University of Birmingham |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://etheses.bham.ac.uk//id/eprint/7603/ |
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