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

该项目将使用蛋白质工程和化学偶联方法来创建新的功能化载铁蛋白（AFt）亚基组合，这些亚基已被定制用于过度表达人表皮生长因子受体（HERx/EGFRs/ErbBx）的肿瘤（乳腺癌、胃癌、卵巢癌）。这些将通过与具有独特化学特征的分子的附着获得特定的肿瘤靶向能力，这些分子对人类表皮生长因子受体具有非常高的亲和力。靶向配体将使用位点特异性非天然氨基酸诱变和随后的生物正交“点击”化学反应以及高度肽序列选择性酶介导的偶联反应的组合来引入。工程纳米载体的自组装能力将用于封装一系列药物分子，该项目将利用荧光和一系列光谱方法研究工程载铁蛋白释放这些药物的机制和速率。纳米载体将使用一系列技术进行全面表征，包括TEM， DLS， MS，共聚焦荧光显微镜。该提案符合EPSRC医疗保健技术的职责范围，因为它涉及到使用合成生物学和化学方法相结合的新型可持续生物材料纳米载体的创建。这些具有巨大的潜力，作为未来治疗的新纳米材料平台，将通过显着更好的靶向性和减少副作用来提高疗效。所生产的纳米载体将受益于其均质性、可扩展的低成本生产和出色的生物相容性，因为它们来源于人类载铁蛋白，并利用了这种纳米笼的独特结构和材料特性。学生将接受培训，使用一系列先进的化学和合成生物学方法来设计新材料，以及使用一系列物理技术来充分表征所生成的新材料。这将通过在合适的人类细胞系中研究药物释放动力学和纳米载体摄取和降解的实践经验来补充，以允许纳米载体设计的迭代改进。

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.