Establishment of mammalian cell expression systems for the production of essential low cost protein reagents for development of rapid diagnostics

目前的SARS-CoV-2大流行突出表明，需要以使中低收入国家和高收入国家广泛获得这些诊断方法的成本，获得具有所需灵敏度和特异性的快速、可靠的诊断方法。基于蛋白质的诊断开发和制造依赖于关键的基于蛋白质的试剂的可用性，这些试剂是这些测试的关键组件。这样的检测可能需要每年生产数百万到数十亿美元，因此需要安全和充足的可信和低成本的蛋白质试剂供应。例如，对于一个简单的疟疾横向流动测试，每百万次测试需要10-500毫克量的3种不同的蛋白质试剂，预计每年需要4亿次测试。其中一些试剂蛋白必须在培养的哺乳动物细胞系统中正确折叠、组装和修饰，以确保它们在诊断中能够以尽可能高的特异性和敏感性发挥作用。虽然有一些蛋白质试剂表达系统可用于从哺乳动物细胞中生产重组蛋白，但这些系统已经得到改进和开发，主要是为了表达高价值的蛋白质，用作治疗一系列疾病和疾病的药物。获得这种系统的成本高得令人望而却步，这反过来又使使用这些必要的试剂蛋白质来制造必要的大容量诊断存在问题。因此，在获得哺乳动物细胞重组蛋白试剂方面存在着大量未得到满足的需求，其成本使其能够应用于商业上可行的诊断。这里描述的项目将在英国领先的诊断公司莫洛奇内建立技术、诀窍和系统，用于在培养的哺乳动物细胞系统中以足够的数量、质量和成本快速生产重组蛋白试剂，以便能够应用于低成本的快速诊断，以便在发达国家和发展中国家分发。这将确保为诊断的开发和制造提供试剂，并提供快速产生新的基本蛋白质试剂的能力，以响应当前和新出现的需求，并以允许它们应用于诊断的成本。

The current SARS-CoV-2 pandemic has highlighted the need for access to rapidly produced, reliable diagnostics with the required sensitivity and specificity at a cost that makes these widely accessible to lower and middle income (LMICs) as well as higher income countries. Protein based diagnostic development and manufacture is reliant upon the availability of key protein based reagents that are critical components of these tests. Such tests may need to be manufactured in their millions to billions a year and thus a secure and sufficient supply of authentic and low cost protein reagents is required. For example, for a simple Malaria lateral flow test, 3 different protein reagents are required in 10-500 mg quantities per million tests with an anticipated need of 400 million tests per annum. Some of these reagent proteins must be made in cultured mammalian cell systems to be correctly folded, assembled and modified in order to ensure they can function with the highest possible specificity and sensitivity in diagnostics. Although there are some protein reagent expression systems for production of recombinant proteins from mammalian cells commercially available, these have been refined and developed largely for the expression of high-value proteins to use as drugs to treat a range of diseases and conditions. Access to such systems involves prohibitively high costs that in turn make the use of these essential reagent proteins to manufacture diagnostics in the necessary high volume problematic. There is thus a massively unmet need in terms of access to mammalian cell recombinant protein reagents at a cost that allows their application into commercially viable diagnostics.The project described here will establish technologies, knowhow and systems within a leading UK diagnostics company, Mologic, for the rapid production of recombinant protein reagents in cultured mammalian cell systems, at sufficient amounts, quality and cost to be viable for application into low cost rapid diagnostics for distribution in both developed and developing countries. This will secure reagent provision for development and manufacture of diagnostics and also provide the ability to generate new essential protein reagents swiftly in response to current and emerging needs at a cost that allows their application into diagnostics.