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结合方法来创建新的功能性载铁蛋白(AFT)亚单位的组合，这些亚单位是针对过表达人类表皮生长因子受体(Herx/EGFRs/ErbBx)的肿瘤(乳腺癌、胃癌、卵巢癌)量身定做的。它们将通过附着具有独特化学特征的分子获得特定的肿瘤靶向能力，这些分子与人类表皮生长因子受体具有非常高的亲和力。靶向配体的引入将结合使用定点特定的非天然氨基酸突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的酶介导的偶联反应。工程纳米载体的自组装能力将被用来包裹一系列药物分子，该项目将使用荧光和一系列光谱方法研究工程载铁蛋白释放这些分子的机制和速度。这些纳米载体将使用一系列技术进行全面表征，包括透射电子显微镜、DLS、MS、共聚焦荧光显微镜。这项建议符合EPSRC医疗技术的范围，因为它涉及创建使用合成生物学和化学方法相结合的新的可持续生物材料纳米载体。这些具有巨大潜力的新纳米材料平台将用于未来的治疗，将通过显著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均质，具有可扩展的低成本生产和良好的生物兼容性，因为它们来自人类载铁蛋白，并利用这种纳米囊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产生的新材料。此外，在研究药物释放动力学和纳米载体在合适的人类细胞系中的摄取和降解方面的实际经验将补充这一点，以便反复改进纳米载体的设计。

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.