Cell Based Assay for Tetanus Vaccine and Antitoxin Production

破伤风疫苗和抗毒素生产的细胞测定

• 批准号: NC/Y000978/1
• 负责人: Andrew Peden
• 金额: $ 43万
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=NC%2FY000978%2F1
• 关键词: Cell; Based; Assay; Tetanus; Vaccine

Tetanus toxin is a member of the Clostridial Toxin family which also includes Botulinum Toxins. This family comprises some of the most potent toxins in existence. They also are used in a wide number of medical applications. Tetanus toxin is chemically treated to make Tetanus Toxoid, which is a non-harmful derivative of Tetanus that is used in Tetanus Vaccine. This vaccine is a WHO Essential Global Medicine. Indeed, it forms the backbone of immunisation schedules across the globe with 83% of the global population currently vaccinated to date. As the bacteria that causes tetanus is present in the soil, it can never be eradicated and therefore the requirement for vaccine will be continuous and expanding along with the global human and livestock populations. As part of the quality control process, tetanus-based products must be tested in animals for potency and, in the case of toxoid, for traces of remaining toxicity. Animal testing is associated with considerable suffering to large numbers of animals and there is an urgent and long recognised need to replace these tests. Previously, we have genetically engineered a cell-line to be highly sensitive to the closely related Botulinum/B. We then developed a robust antibody-based assay which incorporates a luminescent reporter readout for ease of detection of Botulinum/B activity. The assay can detect the very tiny amounts of toxin necessary for potency testing/quality control purposes. Now, we have adapted our technology to be acutely sensitive to Tetanus Toxin and generated state of the art recombinant monoclonal antibodies that can be readily scaled up for global adoption of our technology. In collaboration with the MHRA, we will fully validate our assay for Tetanus Toxin, Antitoxin and importantly for quality control of commercial Toxoids from Vaccine Producers. This project will lay a solid foundation to then immediately translate the technology to end-users for in-house evaluation and ultimately adoption. We will also optimise and "test to destruction" the tolerance of our assay.Many elements of the mechanism of action of tetanus toxin remain to be elucidated owing to the difficulty in studying the specific populations of cells that are sensitive to tetanus in vivo. Our unique portfolio of tools together with recent advances in RNA-sequencing technologies places us in an excellent position to answer these longstanding questions. We will use our engineered cell lines that can be rendered sensitive to tetanus through chemical differentiation, to probe the molecular substrates underlying tetanus sensitivity.

破伤风毒素是梭状芽胞杆菌毒素家族的一员，该家族还包括肉毒毒素。这个家族含有一些现存的最有效的毒素。它们也被广泛应用于医疗领域。破伤风毒素经过化学处理后生成破伤风类毒素，它是破伤风疫苗中使用的一种无害的破伤风衍生物。该疫苗是世卫组织的基本全球药物。事实上，它构成了全球免疫接种计划的支柱，到目前为止，全球有83%的人口接种了疫苗。由于导致破伤风的细菌存在于土壤中，它永远不可能被根除，因此对疫苗的需求将随着全球人畜人口的增加而不断扩大。作为质量控制过程的一部分，基于破伤风的产品必须在动物身上进行效力测试，如果是类毒素，则必须测试残留毒性的痕迹。动物测试与大量动物遭受的痛苦有关，人们迫切需要取代这些测试，这是人们早就认识到的。此前，我们通过基因工程设计了一种对密切相关的肉毒杆菌/B高度敏感的细胞系。然后，我们开发了一种强大的基于抗体的检测方法，该方法结合了发光报告读数，便于检测肉毒杆菌/B的活性。该分析可以检测出效力测试/质量控制所需的非常微量的毒素。现在，我们已经调整了我们的技术，使其对破伤风毒素非常敏感，并产生了最先进的重组单抗，可以很容易地扩大规模，以便在全球采用我们的技术。在与MHRA的合作下，我们将全面验证我们的破伤风毒素、抗毒素检测，重要的是对疫苗生产商的商业类毒素进行质量控制。该项目将奠定坚实的基础，然后立即将该技术转化为最终用户进行内部评估并最终采用。我们还将优化和“测试到破坏”我们的测试的耐受性。由于在体内研究对破伤风敏感的特定细胞群的难度，破伤风毒素作用机制的许多元素仍有待阐明。我们独特的工具组合，加上RNA测序技术的最新进展，使我们处于回答这些长期存在的问题的绝佳位置。我们将使用我们的工程细胞系，通过化学分化使其对破伤风敏感，以探索破伤风敏感性的分子底物。