Integrating living analytics into biomanufacturing processes

细胞越来越多地被用于制造各种各样的药物、可再生塑料、化学品和其他消费品。了解生产过程中细胞的行为是优化生产和确保产品良率和质量的必要条件。目前，这是通过使用基于化学的成熟技术来测量几个因素来完成的。然而，生命系统天生具有感知环境并对变化做出相应反应的能力。这些机制比目前使用的化学方法具有更好的敏感性和特异性。在过去的几年里，科学家们已经开始利用这些自然系统来开发一种叫做生物传感器的分析技术。生物传感器可用于检测环境中感兴趣的分子的存在或缺失。它们也可以用来估计感兴趣的分子（分析物）的浓度。对于生物制造过程来说，后一种类型的生物传感器很有趣，因为它可以精确测量关键因素的浓度，这些因素对于最大化基于细胞的制造系统的产量和生产率非常重要。本提案的目的是开发一个在生物制造过程中使用活体生物传感器的框架。研究的重点是找出最好的方法，让生物传感器和产生细胞一起生长，使两种细胞都能存活并正常运作。为此，我们将尝试许多不同的物理分离方法，包括膜分离，将生物传感器捕获在聚合物中，或将生物传感器捕获在脂质泡中。我们还将尝试对生物传感器细胞进行基因改造，使其粘附在培养容器的壁上，或者使非生命颗粒发芽，这些颗粒仍然可以作为生物传感器，但不能再生长。每一种策略都将使用我们已经开发的生物传感器进行测试，该传感器用于测量来自哺乳动物细胞的分析物的浓度，这些细胞也会产生蛋白质药物。我们希望找到一种条件，使生物传感器不会过度生长在哺乳动物细胞中，但仍然可以感知和响应分析物浓度的变化。一旦活体分析的基本原理被证明，我们希望这将改变未来测量细胞行为的方式，并在制造过程中更好地了解细胞。希望有一天，这将导致生物制造系统的产品产量更高，最终将降低消费者的成本。

Cells are increasingly being used to manufacture a wide variety medicines, renewable plastics, chemicals, and other consumer products. Understanding the behaviour of cells during the manufacturing process is necessary to optimise production and to make sure that product yield and quality are high. Currently, this is done by measuring a few factors using established technologies based on chemistry. However, living systems have naturally have the ability to sense their environment and respond to changes accordingly. These mechanisms have better sensitivity and specificity than the chemical methods currently used. In the past few years, scientists have begun to harness these natural systems to develop analytical technology called biosensors. Biosensors can be used to detect the presence or absence of a molecule of interest in the environment. They also can be used to estimate the concentration of a molecule of interest (analyte). It is the latter type of biosensor that is interesting for biological manufacturing processes because it allows the accurate measurement of concentrations of key factors that are important for maximising the yield and productivity of cell-based manufacturing systems.The aim of this proposal is to develop a framework for using living biosensors in biomanufacturing processes. The focus in on identifying the best way to grow the biosensor and the producing cell together in such away that both types of cells survive and function correctly. For this, we will try a number of different physical separation methods including membrane separation, trapping the biosensor in a polymer, or trapping the biosensor in a bubble of lipid. We will also try to genetically modify the biosensor cell to cause it to stick to the wall of the culture vessel or to bud off non-living particles that can still behave as biosensors, but can no longer grow.Each of these strategies will be tested using a biosensor we have already developed to measure the concentration of an analyte coming from mammalian cells that are also producing a protein drug. We want to find conditions where the biosensor does not overgrow the mammalian cell, but still can sense and respond to changes in analyte concentration. Once the basic principle of a living analytic has been demonstrated, we hope that this will change the way that measurements of cellular behaviour are done in the future and lead to better understanding of cells during the manufacturing process. Hopefully one day this will lead to higher yields of products from biological manufacturing systems, which ultimately will decrease the costs to consumers.