Precision Glycoengineering of an HCV Envelope-Based Nanoparticle Vaccine

HCV 包膜纳米颗粒疫苗的精密糖工程

• 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
• 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

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.

摘要 在全球范围内，超过7100万人感染丙型肝炎病毒（HCV），其中100万至200万人是新感染者。 每年都有感染发生。这一重大健康问题需要开发有效的疫苗。 由于丙型肝炎病毒迅速积累突变，疫苗必须引起广泛中和抗体的产生 （bnAbs）。病毒包膜E1E2糖蛋白是中和的天然靶标。 抗体的然而，生产包膜蛋白如E1E2的两个主要挑战如下。第一、 随着候选蛋白质的发展，获得具有稳定的、可再现的、均质的、 这些糖型显示出产生有效和广泛中和抗体应答的高潜力。这 确保最佳效力的疫苗在临床前和临床试验中的各批次之间具有可比的效力，并且 使其更容易在生产过程中保持聚糖谱。然而，宿主细胞之间的差异，甚至 克隆，可导致糖基化的实质性变化。第二，在鉴定出提供糖基化的糖型时， 由于需要广泛中和的抗体应答，可能难以获得有效的表达宿主， 经济地生产功能形式的疫苗亚单位蛋白。我们在这里应对这些挑战 通过在一组中产生良好表征的新型天然样分泌型E1E2（sE1E2）复合物， 糖工程化的哺乳动物细胞系，以获得具有可预测和确定的 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， 来自用糖工程化CHO细胞系中产生的sE1E2免疫的动物的多克隆血清，野生型CHO 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 并帮助建立一个生产有效泛HCV疫苗的平台细胞系。