Building and commercialisation of a new mammalian cell factory platform

新哺乳动物细胞工厂平台的建设和商业化

• 批准号: 2617838
• 金额: --
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2617838
• 关键词: Building; commercialisation; new; mammalian; cell

Currently industry standard cell factory platforms for manufacture of secreted therapeutic proteins are based around variants of CHO cells, where variants are either natural or engineered auxotroph's for key intermediates in the synthesis of amino acids or nucleotides, essential to support cell growth. Current state-of-the-art recombinant protein production uses genetic vectors which co-express a desired recombinant gene plus a selection marker, using an appropriate CHO cell variant. Only those cells which take up the co-expression vector can be selected on medium that lacks added glutamine (GS) or nucleosides (DHFR). Both systems have the potential for amplification with small molecular weight inhibitors - methionine sulphoxamine (MSX) for GS and methotrexate (MTX) for DHFR. Inclusion of inhibitors in the selection process enables the isolation of cells with higher copies of GS or DHFR which, being genetically linked to the co-expressed recombinant gene, leads to higher expression of the recombinant gene. Consequently, production per cell can be extensively increased and results in productivity suitable for commercial use. Metabolomics studies with CHO cells in the Dickson laboratory have identified a metabolic target that, in a manner analogous to (but distinct from) that of GS, offers a powerful co-expression genetic vector selection system, amenable to knockdown (again analogous to the GS Xceed1 system, developed by Lonza). Several small molecular weight chemicals are potent inhibitors of the metabolic target (analogous to MSX), offering the potential to selection of amplified gene copy. The identified metabolic target is fundamental to cell growth, is in a critical metabolic pathway and, unlike GS, cannot be bypassed by other nutrients such as asparagine. It is also cell-agnostic for potential cell host application. We hypothesise that this platform provides huge potential for synthetic interventions in terms of vector development and host cell line engineering, offering an alternative and improved metabolic selection system to produce therapeutic proteins. This metabolic target and associated gene constitute novel intellectual property. Once exemplified it is intended that the technology would be patented, and that this patent, the materials generated (such as cell lines, vectors and media) would support future commercialisation The PhD student will be responsible for the technical development of the platform, generation of novel cell lines, cell culture medium and other components with design and application of synthetic biology constructs, preparation of knockout cell lines and cell culture components such as medium. Once developed these key components will enable a proof-of-concept evaluation of the platform using a relevant industrial molecule. Production of proteins is of key importance for both therapeutic and nontherapeutic proteins. With global annual sales of therapeutic proteins of >£200Bn, the biomanufacturing sector is an economic leader for the UK. The platform developed in this PhD would be offered as a potential competitor to the technology already reaping commercial dividends for companies such as Lonza, Fuji and Horizon. Currently the lack of freedom to operate and the high cost of entry makes manufacturing expensive and limits the emergence of new companies with innovative products, lacking access to the required technology. A key aim of this project is to develop an innovative, industry-leading, open-source expression platform with significantly flexible license terms, thereby driving significant industry uptake. Revenues would be generated through sales of platform components, technical support and partnerships. The availability of expression platforms un-encumbered by stringent licensing fees would have great attraction to SME's and academic innovators. This expression system has completed an initial proof of concept project undertaken between Pathway Biopharma and an MSc student in the Dickson lab.

目前，用于生产分泌型治疗性蛋白的行业标准细胞工厂平台是基于CHO细胞的变体，其中变体是天然或工程改造的营养缺陷型，用于合成氨基酸或核苷酸的关键中间体，这对支持细胞生长至关重要。目前最先进的重组蛋白生产使用遗传载体，该遗传载体共表达所需的重组基因，外加一个选择标记，使用适当的CHO细胞变体。只有那些携带共表达载体的细胞才能在没有添加谷氨酰胺(GS)或核苷(DHFR)的培养基上被选择。这两个系统都有可能用小分子量抑制剂-蛋氨酸亚磺胺(MSX)用于GS和甲氨蝶呤(MTX)用于DHFR。在选择过程中加入抑制剂可以分离出具有更高拷贝的GS或DHFR的细胞，这些拷贝与共表达的重组基因在基因上相连，导致重组基因的更高表达。因此，每个电池的产量可以得到广泛的提高，从而产生适合商业使用的生产率。在Dickson实验室用CHO细胞进行的代谢组学研究已经确定了一个代谢靶点，它以一种类似(但不同于)GS的方式提供了一个强大的共表达遗传载体选择系统，易于敲除(同样类似于由Lonza开发的GS Xceed1系统)。几种小分子化学物质是代谢靶标(类似于MSX)的有效抑制剂，为扩增基因拷贝的选择提供了可能性。已确定的代谢靶点是细胞生长的基础，处于关键的代谢途径中，与GS不同，不能被天冬酰胺等其他营养物质绕过。对于潜在的细胞宿主应用，它也是细胞不可知的。我们假设，该平台在载体开发和宿主细胞系工程方面为合成干预提供了巨大的潜力，提供了一种替代的和改进的代谢选择系统来生产治疗性蛋白质。这种代谢靶点和相关基因构成了新的知识产权。一旦被例证，该技术将获得专利，并且这项专利将支持未来的商业化。博士生将负责平台的技术开发，生成新的细胞系、细胞培养液和其他组件，设计和应用合成生物学结构，准备敲除细胞系和细胞培养组件，如培养液。一旦开发出来，这些关键组件将使用相关的工业分子对平台进行概念验证评估。蛋白质的生产对于治疗性和非治疗性蛋白质都是至关重要的。全球治疗用蛋白质年销售额达2000亿英镑，生物制造行业是英国的经济领头羊。该博士所开发的平台将成为龙沙、富士和Horizon等公司已经收获商业红利的技术的潜在竞争对手。目前，缺乏经营自由和高昂的进入成本使制造成本高昂，并限制了新公司的出现，这些公司拥有创新的产品，无法获得所需的技术。该项目的一个关键目标是开发一个创新的、行业领先的、开放源码的表达平台，并具有相当灵活的许可条款，从而推动行业的显著采用。收入将通过销售平台组件、技术支持和伙伴关系产生。不受严格许可费制约的表达平台的可获得性将对中小企业和学术创新者具有巨大的吸引力。这个表达系统已经完成了Path Biophma和Dickson实验室的一名理科学生之间进行的初步概念验证项目。