Monoclonal antibodies targeting the fungal cell surface: next generation antifungal therapeutics

念珠菌属和曲霉属的真菌病原体可引起危及生命的侵袭性真菌感染，这些感染可能是抗真菌治疗所不能解决的。有一个明确的临床需要新的抗真菌治疗。真菌细胞壁含有毒力因子，并介导与宿主的相互作用。在靶向方法中，我们设计了重组单克隆抗体，其识别真菌病原体的细胞壁蛋白。这些抗体在准确诊断和治疗真菌感染方面具有巨大的潜力。这个博士项目将使用异源表达的细胞表面蛋白，短肽序列和来自不同临床分离株的真菌细胞来广泛研究我们的先导抗体的结合相互作用。在邓迪大学，货车Aalten实验室已经阐明了几种真菌特异性细胞表面蛋白的结构，并将提供真菌糖蛋白表达方面的专业知识，这些糖蛋白将用于使用生化测定（ELISA）和表面等离子体共振（SPR）表征抗体结合。此外，他们还可以提供蛋白质晶体学方面的专业知识，以引发抗体与真菌靶标的结合，并通过计算机构建这些抗体和靶蛋白的3D结构，以通过界面预测和分子对接研究大分子相互作用。该学生还将在由Soumya Palliyil博士领导的苏格兰生物制品设施接受抗体工程方面的高级培训。利用蛋白质工程的最新工具并结合重组抗体开发的专业知识，该项目还将探索通过将抗菌剂与选定的先导抗体进行生物化学偶联来产生现有抗体的双特异性形式的可行性，例如在同一抗体分子上产生两个不同的抗原结合臂或有效的抗体药物缀合物（ADC），以增强病原体杀灭。

Fungal pathogens of Candida and Aspergillus species cause life-threatening invasive fungal infections that can be recalcitrant to antifungal therapy. There is a clear clinical need for novel antifungal therapies. The fungal cell wall contains virulence factors and mediates interactions with the host. In a targeted approach we have engineered recombinant monoclonal antibodies that recognise cell wall proteins of fungal pathogens. These antibodies have huge potential to accurately diagnose and treat fungal infections. This PhD project will extensively characterise the binding interactions of our lead antibodies using heterologously expressed cell surface proteins, short peptide sequences, and fungal cells from different clinical isolates. At the University of Dundee, the Van Aalten lab has elucidated the structure of several fungal-specific cell surface proteins and will provide expertise in the expression of fungal glycoproteins that will be utilised for the characterisation of antibody binding using biochemical assays (ELISAs) and Surface Plasmon Resonance (SPR). In addition, they can provide expertise in protein crystallography to elicit antibody binding to the fungal targets and in silico construction of the 3D structure of these antibodies and target proteins to study the macromolecular interactions through interface predictions and molecular docking. The student will also receive advanced training in antibody engineering at the Scottish Biologics Facility, headed by Dr Soumya Palliyil. Using the latest tools in protein engineering and combining the expertise of recombinant antibody development, this project will also explore the feasibility of generating bispecific formats of the existing antibodies e.g. two different antigen binding arms on the same antibody molecule or potent antibody drug conjugates (ADCs) by biochemically coupling an antimicrobial agent to the selected lead antibody for enhanced pathogen killing.