Characterization of post-transcriptional constraints that determine rP yield during bioprocessing in mammalian cells

当我们的身体识别出外来“入侵者”时，我们都有一种内在的防御机制来应对感染。一种称为B细胞的细胞通过转变为抗体产生细胞来对感染作出反应。抗体是一种蛋白质，其作用是攻击并摧毁外来入侵者，通过清除外来因子来清除感染。由于抗体是我们身体对疾病的天然防御，许多新的抗体类型的药物正在开发，以帮助治疗一些人类疾病，如癌症。这些抗体通常由在规定条件下保存在培养液中的细胞产生。问题是这些抗体必须是特殊形状的，否则它们就不起作用。用于产生这种抗体的细胞有一套非常复杂的机制来制造抗体，并将它们的成分组合成正确的形状。当预期细胞不会产生大量抗体时，这种方法非常有效。然而，当我们试图生产更多的产品时，我们用来制造抗体的细胞在生产这些药物方面的效率要低得多。因此，我们无法生产足够的这些药物，因此对它们的成本和需求很高，例如乳腺癌药物赫赛汀。问题的根源在于，当细胞被要求产生更多的蛋白质来满足我们的需求时，这种机制再也无法科普/细胞有时会死亡，或者不能产生正确形状的抗体，没有任何临床用途。人们普遍认为，随着进一步开发基于抗体的药物，这个问题将变得更加严重。这里提出的研究将调查制造蛋白质的细胞机制如何工作，并检查是否以及以何种方式可以操纵它来产生更多的抗体。我们想确定这个机器的不同部分在制造抗体方面受到限制，然后研究这些部分如何共同工作，最终产生感兴趣的抗体。目前还不知道这是否可能，并且对目前用于生产抗体的哺乳动物细胞中的过程知之甚少。我们将采用最先进的新技术和方法的组合，以逐步的方式拆开哺乳动物细胞中的抗体组装线，然后使用收集的信息来确定组装过程中每个步骤之间的关系。最终，这将使细胞的操作能够改变装配线中每个步骤的平衡，以降低成本和更高质量生产更多的靶抗体药物。如上所述，这是极其重要的，因为预计随着越来越多的蛋白质“药物”的开发，我们将缺乏生产足够大的量以满足大多数人对这些新药的需求的能力，而不是那些能够负担得起目前仍然非常昂贵但非常有效的药物的人。

We all have an in-built defence mechanism to respond to infection when our body recognises a foreign 'invader'. A type of cell known as a B cell responds to infection by changing into an antibody-producing cell. Antibodies are proteins that work by attacking the foreign invader and destroying it, clearing infection by removing the foreign agent. Because antibodies are our body's natural defence against disease, many new antibody type drugs are being developed to help treat a number of human diseases such as cancer. These antibodies are usually produced by cells kept in a culturing solution under defined conditions. The problem is that these antibodies must be in a special shape; otherwise they do not work. The cells used to produce such antibodies have a very complex set of machinery to make the antibodies and put their components together into the right shape. This works very well when the cell is not expected to make much of the antibody in question. However, the cells we use to make antibodies are much less efficient at producing these drugs when we try and produce more of the product. As a result, we are not able to produce enough of these drugs and the cost and demand for them is therefore high e.g. the breast cancer drug Herceptin. The root of the problem is that when cells are asked to produce much more protein to meet our needs the machinery can no longer cope / the cells sometimes die or else don't produce antibodies of the right shape, of no clinical use whatsoever. It is largely agreed that this problem will become even serious as further antibody-based drugs are developed. The research proposed here will investigate how the cell machinery for making proteins works and examine whether, and in what ways, it can be manipulated to produce more antibody. We want to determine the different parts of this machinery that are limiting in terms of making the antibody, and then investigate how these parts work together to ultimately produce the antibody of interest. At present it is unknown if this is possible, and the process is poorly understood in the mammalian cells presently used to produce antibodies. We will employ a combination of new state-of-the-art technologies and approaches to take apart the antibody assembly line in mammalian cells in a step-wise manner, and then using the information gathered will determine the relationship between each step of the assembly process. Ultimately this should enable the manipulation of cells to change the balance of each step in the assembly line to produce more of the target antibody drug at reduced cost and higher quality. As stated above, this is extremely important as it is expected that with an increasing number of protein 'drugs' being developed we will lack the capability of producing large enough amounts to meet the required demand for these new drugs for the majority, as opposed to for those who can afford what must currently remain prohibitively expensive, but very effective, medicines.