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

寡糖-融合蛋白缀合物的合成方法及其抗原性增强

• 批准号: 10078593
• 负责人: Steven Sucheck
• 金额: $ 43.67万
• 依托单位: UNIVERSITY OF TOLEDO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10078593
• 关键词: Address; Adjuvant; Animal Model; Antibodies; Antibody Response; Antibody titer measurement; Antigen Targeting; Antigen-Presenting Cells; Antigens; Antimicrobial Resistance; Bacteria; Bacterial Antigens; C-terminal; CD8-Positive T-Lymphocytes; CD8B1 gene; Carbohydrates; Cell Culture Techniques; Cessation of life; Chemicals; Chemistry; Chimeric Proteins; Collaborations; Complex; Cysteine; Cytotoxic T-Lymphocytes; Development; Dose; ESKAPE pathogens; Economics; Engineering; Epitopes; Escherichia coli; Extracellular Domain; Family; Fc domain; Glycoconjugates; Goals; Health; Helper-Inducer T-Lymphocyte; Human; Hybrids; IgG3; Immune response; Immune system; Immunization; Immunology; Inbred Mouse; Lipids; Lipopolysaccharides; Liposomes; Mediating; Medicine; Membrane Proteins; Methods; Molecular; Molecular Biology; Mus; Oligosaccharides; OprF protein; Organic Chemistry; Pathogenesis; Peptides; Play; Polymers; Polysaccharides; Production; Proteins; Pseudomonas aeruginosa; Publishing; Rhamnose; Role; Route; Serotyping; Site; T-Lymphocyte; Tertiary Protein Structure; Tumor Antigens; Vaccine Antigen; Vaccines; Viral Antigens; Work; base; cost; cytokine; design; experimental study; humanized mouse; immunogenicity; improved; in vivo; investigator training; member; mouse model; novel; novel strategies; pathogen; receptor; recruit; response; single molecule; synthetic biology; vaccine candidate

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.

项目摘要 碳水化合物已被用于世界上一些最有效的糖共轭化合物 疫苗。然而，传统的糖偶联物是通过低聚糖分离得到的 未能生产出针对ESKAPE病原体假单胞菌的有效疫苗 铜绿假单胞菌。ESKAPE病原体是已经获得广泛的 抗菌素耐药性(AMR)和正在变得越来越困难，在一些 无法治疗的病例。迫切需要确定新的方法来解决 控制AMR，预计每年导致多达1000万人死亡，并造成成本 到2050年，累计经济损失将达到100万亿美元。此应用程序 通过使用一种强大的基于可溶性聚合物的合成方法来解决AMR 用化学方法合成非血清型低聚糖表位 铜绿假单胞菌的脂多糖。此外，我们还开发了健壮的 将这些人工合成的低聚糖结构域与 基于合成生物学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偶联物与脂类定位偶联的方法 佐剂形成定义明确的单分子脂糖蛋白。这些材料将是 用于确定低聚糖和蛋白质在体内的有益作用 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。这些抗原的设计是为了克服 对此产生保护性免疫反应的已知障碍 细菌，并提供对多种细菌菌株的保护。的第二个方面 该计划是对小鼠体内产生的结合物进行系统研究，这将导致 增强人类疫苗免疫原性的新方法。这种方法采取了 人体内自然产生的抗体可参与增强 免疫系统对疫苗的识别。这项建议充分利用了 受过有机化学、免疫学培训的研究人员和 发病的分子机制。