Developing novel label-free technologies for studying drug molecules and their ability to induce protein complex formation in cells

在模型体外系统和活细胞中实时成像药物疗效对于制药工业中新药物分子的开发至关重要。传统上，药物被化学修饰，以便于在显微镜下使用例如荧光观察和作用模式，但这是有问题的，因为这些修饰导致药物疗效的变化。因此，开发新的成像技术，允许对药物进行无标记、无光毒性的成像，对于解锁药物发现管道至关重要。这种无标记成像的应用将包括研究药物分子触发蛋白质复合体形成的能力。这项研究将通过开发利用折射率变化的新型无标记成像技术(全息断层成像)来直接解决这一瓶颈。这项令人兴奋的技术将与模型膜和合成生物细胞相结合，其中嵌入了感兴趣的蛋白质复合体。通过将这些生物膜暴露于候选药物，将有可能动态监测蛋白质复合体的形成。这些合成细胞将被排列，以产生高通量屏幕，随后的研究将着眼于3-D打印细胞和芯片上器官平台的复杂形成。所有结果将通过荧光成像和光谱学进行验证，包括单分子方法，如荧光相关光谱(FCS)。这一令人兴奋的项目将使成功的学生申请者在微流体、无标记成像、3D生物打印、细胞培养、合成细胞构建和药物发现科学方面获得丰富的经验。

Real time imaging of drug efficacy in model in-vitro systems and living cells is vital to the development of novel drug molecules in the pharmaceutical industry. Traditionally drugs are chemically modified to facilitate their observation and mode of action under a microscope using e.g. fluorescence but this is problematic as these modifications result in changes in drug efficacy. The development of novel imaging techniques that allows for label-free imaging of drugs with no phototoxicity is therefore vital to unlocking drug discovery pipelines. Applications of such label free imaging would include studying the ability of drug molecules to trigger protein complex formation. This studentship will directly address this bottleneck through the development of novel label free imaging techniques (holotomography) that exploit changes in refractive indices. This exciting technology will be coupled with model membranes and synthetic biological cells into which protein complexes of interest have been embedded. By exposing these biological membranes to drug candidates it will then be possible to dynamically monitor the formation of protein complexes. These synthetic cells will be arrayed so as to generate high-throughput screens, with subsequent studies looking at complex formation in 3-D printed cells and organ on a chip platforms. All results will be validated with fluorescence imaging and spectroscopy, including single molecule approaches such as fluorescence correlation spectroscopy (FCS).This exciting project will enable the successful student applicant to gain extensive experience of microfluidics, label-free imaging, 3D-bioprinting, cell culture, synthetic cell construction and drug discovery science.