Precision Glycoengineering of an HCV Envelope-Based Nanoparticle Vaccine

HCV 包膜纳米颗粒疫苗的精密糖工程

基本信息

批准号：
10759994
负责人：
Stephen Kevin Horrigan
金额：
$ 30万
依托单位：
NEUIMMUNE BIOLOGICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-17 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10759994
关键词：
2019-nCoV Address Animal Model Animals Antibodies Antibody Response Antigen Presentation Antigens B-Lymphocytes Binding Biochemical Biological Assay Biophysics Cell Line Cells Chinese Hamster Ovary Cell Clinical Trials Complex Coupled Coupling Data Development Ebola Engineering Ensure Enzyme-Linked Immunosorbent Assay Epitopes Evaluation Formulation Genotype Glycoengineering Glycoproteins HIV Health Hepatitis C Vaccine Hepatitis C virus Heterogeneity Immune Immune Evasion Immune response Immune system Immunize Immunologic Tests Immunologics Infection Liver Location Mammalian Cell Measures Medical Mus Mutation Persons Pharmacologic Substance Phase Polysaccharides Population Process Production Property Protein Isoforms Protein Subunits Proteins Recombinant Proteins Reproducibility Research Series Serum Side Site Structure Subunit Vaccines Surface System Systems Biology Test Result Vaccine Design Vaccine Production Vaccines Validation Variant Viral Viral Antigens Viral Proteins Virus Work analysis pipeline analytical method cell type comparative efficacy design env Gene Products global health glycosylation immunogenicity improved in vitro testing insight interest manufacture nanoparticle neutralizing antibody novel particle pre-clinical preclinical trial receptor receptor binding response vaccine candidate vaccine development virus envelope

项目摘要

ABSTRACT Globally, more than 71 million people are infected with Hepatitis C virus (HCV), with 1-2 million new infections occurring each year. This major health concern necessitates the development of an effective vaccine. Since HCV rapidly accumulates mutations, vaccines must elicit the production of broadly neutralizing antibodies (bnAbs) in a reproducible fashion. The viral envelope E1E2 glycoprotein is a natural target of neutralizing antibodies. However, two major challenges in production of envelope proteins such as E1E2 are as follows. First, as the candidate protein advances, it will be critical to obtain a product with stable, reproducible, homogenous glycoforms that show high potential for yielding a potent and broadly neutralizing antibody response. This ensures an optimally potent vaccine with comparable efficacy across batches in preclinical and clinical trials, and makes it easier to retain the glycan profile through manufacturing. However, variation across host cells, even clones, can lead to substantial variation in glycosylation. Second, upon identifying a glycoform that provides the desired broadly neutralizing antibody response, it can be difficult to obtain an effective expression host that can economically produce the vaccine subunit proteins in a functional form. Here we are addressing these challenges by producing a well-characterized novel native-like secreted E1E2 (sE1E2) complex in a panel of glycoengineered mammalian cell lines to obtain more homogeneous glycosylation with predictable and defined structures. After production of preclinical material of all glycoforms, the most effective glycoforms will be identified by rigorous bioanalytical analysis, coupled to nanoparticles, and subjected to immunological assessment of polyclonal sera from animals immunized with sE1E2 produced in glycoengineered CHO cell lines, wild-type CHO cell lines, and HEK 293 and Huh7 control cell lines. Thus, this proposed research will identify optimal glyforms and help establish a platform cell line for manufacturing an effective pan-HCV vaccine.

抽象的全球有超过 7100 万人感染丙型肝炎病毒 (HCV)，其中新增 1-200 万人感染每年都会发生。这一重大健康问题需要开发有效的疫苗。由于 HCV 会迅速积累突变，因此疫苗必须引发广泛中和抗体的产生（bnAbs）以可重复的方式。病毒包膜 E1E2 糖蛋白是中和的天然靶标抗体。然而，E1E2 等包膜蛋白的生产面临以下两个主要挑战。第一的，随着候选蛋白质的进展，获得稳定、可重复、均质的产品至关重要糖型具有产生有效且广泛中和抗体反应的巨大潜力。这确保在临床前和临床试验中各批次疫苗的效力具有可比性，并且使得在制造过程中更容易保留聚糖特征。然而，宿主细胞之间的差异，甚至克隆，可导致糖基化的显着变化。其次，在鉴定出提供以下功能的糖型后由于需要广泛中和抗体反应，因此很难获得有效的表达宿主经济地生产功能形式的疫苗亚基蛋白。我们正在解决这些挑战通过在一组糖工程哺乳动物细胞系以获得更均一的糖基化，具有可预测和明确的结构。在生产所有糖型的临床前材料后，将鉴定最有效的糖型通过严格的生物分析，与纳米颗粒偶联，并进行免疫学评估来自用 sE1E2 免疫的动物的多克隆血清，在糖工程 CHO 细胞系、野生型 CHO 中产生细胞系，以及 HEK 293 和 Huh7 对照细胞系。因此，这项拟议的研究将确定最佳的糖型并帮助建立用于生产有效的泛丙型肝炎疫苗的平台细胞系。