Beyond Antibodies - Pick 'n' Mix Modified Apoferritin Subunits Harnessing Self-assembly for Targeted Therapy

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This project will use both protein engineering and chemical conjugation methods to allow the creation of a portfolio of new functionalized apoferritin (AFt) subunits that have been tailored to target tumours (breast, gastric, ovarian cancer) that overexpress Human epidermal growth factor receptors (HERx/EGFRs/ErbBx). These will gain specific tumour targeting abilities through the attachment of molecules with unique chemical signatures that have very high affinities for the Human epidermal growth factor receptors. The targeting ligands will be introduced using a combination of site-specific un-natural amino acid mutagenesis and subsequent bio-orthogonal 'click' chemistry reactions as well as highly peptide sequence selective enzyme-mediated conjugation reactions. The self-assembly abilities of the engineered nanocarriers will be used to encapsulate a range of drug molecules and the project will investigate the mechanism and rate of release of these by the engineered apoferritin using fluorescence and a range of spectrometry methods. The nanocarriers will be fully characterized using a range of techniques including TEM, DLS, MS, confocal fluorescence microscopy.This proposal fits the EPSRC Healthcare technologies remit as it involves the creation of new sustainable biomaterial nanocarriers engineered using a combination of synthetic biology and chemical methods. These have significant potential as new nanomaterial platforms for future therapies that will have improved efficacy through significantly better targeting and reduce side effects. The nanocarriers produced will benefit from being homogeneous, having scalable low-cost production and excellent biocompatibility given they are derived from human apoferritin and exploit the unique structural and material properties of this nanocage.The student will be trained in the site specific manipulation of proteins using a range of advanced chemical and synthetic biology methods to engineer new materials as well as in the use of a range of physical techniques to fully characterize the new materials generated. This will be complemented by practical experience in studying the drug release kinetics and nanocarrier uptake and degradation in suitable human cell lines to allow iterative improvement of the nanocarrier design.