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.

关于无项目蛋白蛋白质合成（CFP）具有革新生物制造的潜力。传统的，基于细胞的蛋白质表达工作流程需要细胞系转化才能表达感兴趣的基因，然后将所得菌株培养到可以提取足够产物的有用体积。这些步骤中的每个步骤本身都是一个工程挑战，必须重复并重新优化每种蛋白质。 CFP完全绕过这一点，从细胞的孤立翻译机制中产生蛋白质。在许多参数中表现良好的一个无细胞系统称为爱丽丝（几乎没有活细胞表达），它包含浓缩的蛋白质合成机制，这些蛋白质合成机制源自烟草根细胞培养物，并由Leniobio商业化。已显示它提供了> 3 mg/ml的参考蛋白质产量，并从微层到升升提供了便捷的缩放电位。该真核系统具有产生一系列困难蛋白质的潜力，包括膜蛋白和病毒样颗粒蛋白。尽管取得了这些早期的地标成功，但在爱丽丝系统中，还有更多的理解和工程，以进一步实现按需生物制造制造。在这个项目中，我们将研究爱丽丝的使用和生产，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。爱丽丝成分（例如核糖体，微粒体和/或线粒体）和蛋白质生产反应的分析和表征将采用来理解这些CQA，并最终将这些学习转化为商业制造过程。在此计划中，将成为系统应用的机会和蛋白质疗法/疫苗的量表生产的机会。这将允许将项目的最佳实践发展用于现实世界应用，并通过随后的功能表征匹配缩放蛋白质的生产。目的1）为爱丽丝系统建立关键质量属性（CQA），并设计可用于进程和批处理释放质量控制的分析程序，重点关注整个爱丽丝裂变，翻译机械和天然微粒体。2）2）对与爱尔丽斯的反应进行机械理解，以实现AliCe Recessions的机械理解。研究方法：候选者将发展生物信息学，微生物学，分子生物学，无细胞蛋白质合成，蛋白质纯化，生化/生物物理学方法，显微镜，显微镜，分析和生物处理方法。与EPSRC的策略和研究领域保持一致：该项目与战略优先级“工程和技术领域的前沿”以及“改造健康和医疗保健”以及研究领域的“制造技术”，因为它研究了具有高效率/可靠性/适当精确/灵活性的制造过程，产品和系统。