Development of a novel inducible expression system for the manufacture of therapeutic proteins from CHO cells.

开发一种新型诱导表达系统，用于从 CHO 细胞生产治疗性蛋白质。

• 批准号: 104199
• 金额: $ 17.61万
• 依托单位: SYNPROMICS LTD
• 依托单位国家: 英国
• 项目类别: Collaborative R&D
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=104199
• 关键词: Development; novel; inducible; expression; system

Production of biopharmaceuticals that are toxic or have detrimental effects on the growth of the host cell line is very challenging. To overcome this challenge researchers have developed a number of promoters allowing control of protein expression during the bioproduction process. The primary aim of that research was to enable a 2-step bioprocessing platform to be developed whereby the expression of the gene of interest is switched on after sufficient biomass has been obtained or a gene that has negative effects on DSP or product quality can be silenced. This type of process is particularly attractive for the production of difficult to express or toxic proteins where expression is deleterious to the cell or where host cell protens co-purify with the final product and complicate purification strategies. However, to date these promoters have proven to have insufficient control for the task with leaky expression observed or the need for multiple rounds of transfection and cell line selection to achieve the requisite control. Ideally the activity of those promoters should be inducible/repressible by a stress stimulus (chemical or physical) during the production process and allow tight control over the expression levels to ensure maximum productivity and high quality of the final product. To this end Synpromics has used its proprietary technology to develop new synthetic inducible/repressible systems that have demonstrated exquisite control of gene expression. These systems offer significant advantages over the currently available systems as they are small in size, can be driven from one plasmid, offer fine tuning of gene expression, are driven by physiological or chemical stimulus that are non-toxic and are therefore ideal for improving productivity and minimising costs during bioproduction in CHO cells. Using these novel gene expression control tools Synpromics and Lonza will embark on an 18 month exemplification program to validate the use of these control tools in a GMP environment using well characterized model proteins. Once validated in an industrially relevant situation, these tools will then be used to design a process for a therapeutic construct and a manufacturing process will be developed. In addition to this outcome, the validation of Synpromics's control tools will increase the flexibility of bioproduction and offer the industry new powerful tools with which to increase the number and type of proteins that can be produced from CHO cells.

对宿主细胞系的生长具有毒性或有害影响的生物药物的生产是非常具有挑战性的。为了克服这一挑战，研究人员开发了许多启动子，允许在生物生产过程中控制蛋白质表达。该研究的主要目的是开发一个两步生物处理平台，从而在获得足够的生物量后启动感兴趣基因的表达，或者可以沉默对DSP或产品质量有负面影响的基因。这种类型的方法对于生产难以表达或有毒的蛋白质特别有吸引力，其中表达对细胞有害或其中宿主细胞蛋白质与最终产物共纯化并使纯化策略复杂化。然而，迄今为止，这些启动子已被证明对于观察到的泄漏表达或需要多轮转染和细胞系选择以实现必要的控制的任务具有不足的控制。理想地，这些启动子的活性应该是在生产过程中通过胁迫刺激（化学或物理）诱导/抑制的，并且允许严格控制表达水平以确保最终产品的最大生产率和高质量。为此，Synproximate利用其专有技术开发了新的合成诱导/抑制系统，该系统已证明可以精确控制基因表达。这些系统提供了优于目前可用系统的显著优点，因为它们尺寸小，可以从一个质粒驱动，提供基因表达的微调，由无毒的生理或化学刺激驱动，因此对于在CHO细胞中生物生产期间提高生产率和最小化成本是理想的。使用这些新型基因表达控制工具，Synpromics和Lonza将开始一项为期18个月的示范计划，使用经过充分表征的模型蛋白来验证这些控制工具在GMP环境中的使用。一旦在工业相关情况下得到验证，这些工具将用于设计治疗性构建体的工艺，并开发制造工艺。除了这一结果，Synprostaglandin的控制工具的验证将增加生物生产的灵活性，并为行业提供新的强大工具，以增加CHO细胞生产的蛋白质的数量和类型。