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

基本信息

批准号：
10320364
负责人：
Steven Sucheck
金额：
$ 44.27万
依托单位：
UNIVERSITY OF TOLEDO
依托单位国家：
美国
项目类别：
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/10320364
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 vaccine immunogenicity

项目摘要

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.

项目概要碳水化合物已被用于一些世界上最有效的糖复合物疫苗。然而，通过寡糖分离生产的传统糖复合物未能生产出针对 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传播的病原体抗菌素耐药性（AMR）变得越来越困难，在某些情况下无法治疗的病例。迫切需要找到新的方法控制抗菌素耐药性预计每年会导致多达 1000 万人死亡，且成本高昂到 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基于可溶性聚合物的强大合成方法来解决 AMR 化学合成不依赖于血清型的寡糖表位铜绿假单胞菌的脂多糖（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。我们开发方法进一步将这些糖缀合物与脂质进行位点特异性缀合佐剂形成明确的单分子脂糖蛋白。这些材料将被用于确定寡糖和蛋白质的有益作用激发保护性免疫反应。此外，这项工作还将允许绘制地图保护性寡糖和蛋白质表位。抗原旨在克服针对这种情况的保护性免疫反应发展的已知障碍细菌并提供针对多种细菌菌株的保护。第二个方面该提案是对小鼠体内产生的缀合物进行系统研究，这将导致增强人类疫苗免疫原性的新方法。该方法采取人类天然存在的抗体的优势可以参与增强免疫系统对疫苗的识别。该提案利用了接受过有机化学、免疫学和发病机制的分子机制。