Methods to synthesize oligosaccharide-fusion protein conjugates and enhancement of their antigenicity

项目摘要 碳水化合物已成为世界上一些最有效的糖缀合物 疫苗。然而，通过寡糖分离产生的传统糖缀合物 未能生产针对ESKAPE病原体假单胞菌的有效疫苗 铜绿。ESKAPE病原体是已经获得广泛感染的病原体， 抗生素耐药性（AMR），并变得越来越困难，在一些 无法治疗的病例。迫切需要确定新的方法， 控制AMR，预计每年造成多达1000万人死亡， 到2050年累积的经济损失将达到100万亿美元。本申请 通过使用强大的基于可溶性聚合物的合成方法解决AMR问题， 化学合成非依赖型寡糖表位， 铜绿假单胞菌细菌的脂多糖（LPS）。此外，我们还开发了强大的 将这些合成寡糖结构域与 通过基于合成生物学的方法制备的保护性融合蛋白结构域。我们 开发进一步将这些糖缀合物与脂质位点特异性缀合的方法 佐剂以形成明确的单分子脂糖蛋白。这些材料将 用于确定低聚糖和蛋白质在以下方面的有益作用： 引发保护性免疫反应此外，这项工作将允许绘制 保护性寡糖和蛋白质表位。抗原的设计是为了克服 已知的障碍发展的保护性免疫反应， 细菌，并提供针对多种细菌菌株的保护。的第二方面 该建议是在小鼠中对所得缀合物进行系统研究， 增强疫苗在人体内免疫原性的新方法。该方法需要 人体内天然存在的抗体的优势， 免疫系统对疫苗的识别该提案利用了 在有机化学、免疫学和生物医学方面受过训练的研究人员之间的合作， 发病的分子机制。

Project Summary Carbohydrates have been used in some of the world most effective glycoconjugate vaccines. However, traditional glycoconjuates produced by oligosaccharide isolation have failed to produce effective vaccines against the ESKAPE pathogen Pseudomonas aeruginosa. ESKAPE pathogens are pathogens that have acquired extensive antimicrobial resistance (AMR) and are becoming increasingly difficult and in some cases impossible to treat. There is an urgent need to identity new approaches for controlling AMR which is projected to cause up to 10 million deaths annually and cost 100 trillion dollars in cumulative economic damage by the year 2050. This application addresses AMR by using a powerful soluble polymer-based synthetic method to chemically synthesize serotype-independent oligosaccharide epitopes from the lipopolysaccharide (LPS) of P. aeruginosa bacteria. In addition, we develop robust methods to site-specifically conjugate these synthetic oligosaccharide domains with protective fusion protein domains prepared by synthetic biology-based methods. We develop methods to further site-specifically conjugate these glycoconjugates with lipid adjuvants to form well-defined single molecule lipoglycoproteins. These materials will be used to determine the beneficial role of both the oligosaccharide and the protein in provoking a protective immune response. In addition, the work will allow mapping of the protective oligosaccharide and protein epitopes. The antigens are designed to overcome known obstacles to development of a protective immune response against this bacterium and to provide protection against multiple bacterial strains. A second aspect of the proposal is the systematic study of the resulting conjugates in mice that will lead to new approaches to enhance vaccine immunogenicity in humans. The approach takes advantage of naturally occurring antibodies in humans that can participate in enhancing the recognition of the vaccine by the immune system. This proposal takes advantage of a collaboration between investigators trained in organic chemistry, immunology, and molecular mechanisms of pathogenesis.