Application of metabolomics profiling of recombinant mammalian cells to bioprocess design

许多临床疾病的诊断和治疗依赖于新的治疗药物/复杂蛋白质分子的开发，这些药物/复合蛋白分子需要在培养的哺乳动物细胞中产生。基因工程的进步和对基因表达的理解使人们有可能将特定培养条件下的哺乳动物细胞转变为“工厂”，从而能够收获有价值的治疗蛋白质药物。因此，有可能在培养中的哺乳动物细胞中引入一种基因，该基因将直接合成所需的药物，并允许哺乳动物细胞执行产生治疗性蛋白质所需的所有必要反应。这些方法产生的一些药物是家喻户晓的(如胰岛素和抗血栓药物)，另一些则不是(如抗体)，然而，所有这些都是治疗许多危及生命的临床疾病的关键和必不可少的成分。一个关键问题是，开发和生产足够数量的治疗性蛋白质药物受到细胞培养产生足够产量的能力的限制。因此，投入开发和加工时间以获取足够数量的治疗性蛋白质使这一过程成本高昂，而这些都是昂贵的药物。尽管工业和学术研究人员付出了巨大努力，但我们仍然不能完全了解限制细胞培养系统产量的因素，如果我们要设计增加产量的方法，这些信息将是必不可少的。这些治疗性蛋白质药物中某些药物的单位成本影响到治疗的可获得性，而对于某些“难以制造”的药物，财务影响可能会阻碍该药物的开发上市。这项建议解决了这个问题，并试图使用新的方法来询问我们如何改进哺乳动物细胞在培养中形成治疗性蛋白质的过程。我们的方法是对细胞内限制所需药物形成的事件进行广泛的观察。研究细胞功能的实验方法往往集中在一个特定的方面，或者检查细胞中的功能，而这些功能并不直接确定是否参与了所研究的过程。我们正在建立复杂的模型，这些模型将确定细胞中的许多因素如何相互作用，以定义治疗性蛋白质的生产是如何控制的。然后，我们将测试该模型，以响应我们在细胞中强加的条件，并由此确定是否有一种方法来识别群体中具有理想特征的细胞，从而产生出色的生产，或者我们是否可以将这些功能工程到细胞中。该计划中的研究团队来自不同的研究领域，使相互作用中出现的各种技术得以融合。分析科学家、细胞生物学家、化学工程师和数学模型师的融合产生了一种新的基于团队的方法来解决工业(并最终)与临床相关的问题。该团队将是一个更广泛的科学家网络的一部分(在学术和工业团体中)，他们是工业俱乐部生物处理研究的一部分，该俱乐部是政府/工业倡议，以保持英国在该领域的研究实力。

The diagnosis and treatment of many clinical conditions is dependent on developments of new therapeutic drugs / complex protein molecules that require production by mammalian cells in culture. Advances in genetic engineering and understanding in gene expression has made it possible to turn mammalian cells in defined culture conditions into 'factories' to permit harvest of the valuable therapeutic protein drugs. Hence it is possible to introduce into mammalian cells in culture a gene that will direct synthesise the desired drug and allow the mammalian cells to perform all the necessary reactions required to generate the therapeutic protein. Some of the drugs generated by these approaches are household names (eg insulin and clot-buster drugs) and others are not (eg antibodies), however all are critical and essential components in treatments for many life-threatening clinical conditions. A key problem is that the development and generation of sufficient amounts of therapeutic protein drugs is limited by the capacity to generate sufficient production from the cell cultures. Consequently, the investment of development and process time to harvest sufficient amounts of therapeutic proteins makes the process costly and these are expensive drugs. Despite significant effort by industrial and academic researchers we still do not fully understand the factors that limit production by the cell culture system and such information will be essential if we are to devise means to increase production. The unit cost for certain of these therapeutic protein drugs has implications for availability for treatment and for certain 'difficult-to-make' drugs financial implications may prevent the drug being developed for the market. This proposal addresses this problem and seeks to use novel approaches to ask how we can improve the process of therapeutic protein formation by mammalian cells in culture. Our approach is to take a wide vision of the events within cells that limit formation of the desired drug. Too frequently experimental approaches to study the functions of cells focus on one specific aspect or examine functions in the cell for which there is no direct certainty of involvement in the processes under study. We are building complex models that will determine how the many factors in the cell interact to define how production of the therapeutic protein is controlled. We will then test the model in response to conditions that we impose in cells and, from this, determine if there is a means to identify those cells in a population with desirable characteristics that will give excellent production or if we can engineer these functions into cells. The research team in this programme have come together from distinctive areas of research, allowing a fusion of technologies to emerge from the interactions. The fusion of analytical scientists, cell biologists, chemical engineers and mathematical modellers generates a novel team-based approach to an industrially- (and, ultimately) clinically-relevant issue. The team will be part of a wider network of scientists (in academic and industrial groups) who are part of the Bioprocessing Research for Industry Club, the government/industrial initiative to maintain the UK research strength in this area.