Enabling on-demand biologics manufacturing with ALiCE

通过 ALiCE 实现按需生物制剂制造

• 批准号: 2881247
• 金额: --
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 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=studentship-2881247
• 关键词: Enabling; demand; biologics; manufacturing; ALiCE

About the ProjectCell-free protein synthesis (CFPS) has the potential to revolutionise biomanufacturing. Traditional, cell-based protein expression workflows require cell line transformation to express a gene of interest, followed by culture of the resulting strain to useful volumes where sufficient product can be extracted. Each of these steps is an engineering challenge in and of itself and must be repeated and reoptimized for every protein. CFPS circumvents this entirely, producing protein in a matter of hours from the isolated translational machinery of cells. One cell-free system that performs well across many parameters is called ALiCE (Almost Living Cell Free Expression), it contains the concentrated protein synthesis machinery derived from tobacco root cell cultures and is commercialised by LenioBio. It has been shown to provide reference protein yields of >3 mg/mL and facile scaling potential from microliters through to liters. This eukaryotic system has the potential to produce a range of difficult proteins, including membrane proteins and virus-like particle proteins. Despite these early landmark successes, there is more to understand and engineer within the ALiCE system to further enable on-demand biologics manufacturing. Aims In this project we will investigate the use and production of ALiCE, with an emphasis on identifying Critical Quality Attributes (CQAs) of the system that will inform best practice manufacturing. Analysis and characterisation of ALiCE components (e.g. ribosomes, microsomes and/or mitochondria) and the protein production reaction will be employed to understand these CQAs and ultimately translate these learnings into a commercial manufacturing process.Underpinning this program of research, will be the opportunity for system application and the scaled production of protein therapeutics/vaccine candidates. This would allow the best practice developments of the project to be leveraged for a real-world application, with scaled protein production matched by subsequent functional characterisation. Objectives1) Establish Critical Quality Attributes (CQAs) for the ALiCE system and devise analytical procedures that could be implemented for in-process and batch release quality control, with focus on whole ALiCE lysate, translation machinery and native microsomes.2) Develop a mechanistic understanding of the ALiCE reaction to enable modelling of engineering interventions.3) Apply ALiCE for on-demand production of therapeutics or vaccine candidates combined with physiochemical and functional protein characterisation.Research methods:The candidate will develop skills in bioinformatics, microbiology, molecular biology, cell-free protein synthesis, protein purification, biochemical/biophysical methods, microscopy, analytics, and bioprocessing. Alignment to EPSRC's strategies and research areas:This project is aligned with the Strategic Priorities 'Frontiers in Engineering and Technology' and 'Transforming Health and Healthcare,' as well as the Research Area 'Manufacturing Technologies', because it investigates manufacturing processes, products, and systems that function with high efficiency/reliability and appropriate precision/flexibility.

关于项目无细胞蛋白质合成(CFPS)有可能给生物制造带来革命性的变化。传统的基于细胞的蛋白质表达工作流程需要细胞系转化来表达感兴趣的基因，然后将所产生的菌株培养到有用的体积，从而可以提取足够的产品。这些步骤中的每一步本身都是一个工程挑战，必须针对每种蛋白质重复和重新优化。CFPS完全绕过了这一点，在几个小时内就能从孤立的细胞翻译机制中产生蛋白质。一种在许多参数下表现良好的无细胞系统被称为ALICE(几乎活着的细胞自由表达)，它包含来自烟草根细胞培养物的浓缩蛋白质合成机制，并由LenioBio商业化。它已被证明提供了3毫克/毫升的参考蛋白质产量和从微升到升的简易缩放潜力。这个真核系统有可能产生一系列困难的蛋白质，包括膜蛋白和病毒样颗粒蛋白。尽管取得了这些早期的里程碑式的成功，但在Alice系统中还有更多的东西需要了解和设计，以进一步实现按需生物制剂制造。在这个项目中，我们将调查Alice的使用和生产，重点是确定系统的关键质量属性(CQA)，这将为最佳实践制造提供信息。对ALICE组分(例如核糖体、微生物体和/或线粒体)和蛋白质生产反应的分析和表征将被用来理解这些CQA，并最终将这些知识转化为商业制造过程。在这项研究计划的支持下，将有机会进行系统应用和蛋白质疗法/疫苗候选的规模化生产。这将允许该项目的最佳实践开发被用于现实世界的应用，规模化的蛋白质生产与随后的功能特征相匹配。目标1)为Alice系统建立关键质量属性(CQA)，并设计可用于过程中和批量释放质量控制的分析程序，重点放在整个Alice裂解产物、翻译机器和天然微生物体上。2)从机理上理解Alice反应，以便能够对工程干预进行建模。3)结合理化和功能蛋白质特性，应用Alice按需生产治疗或疫苗候选。研究方法：候选人将发展生物信息学、微生物学、分子生物学、无细胞蛋白质合成、蛋白质纯化、生化/生物物理方法、显微镜、分析和生物处理方面的技能。与EPSRC的战略和研究领域保持一致：该项目与战略重点“工程和技术前沿”、“转型健康和医疗保健”以及研究领域“制造技术”保持一致，因为它研究以高效率/可靠性和适当的精度/灵活性运行的制造过程、产品和系统。