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，需要快速生产具有所需灵敏度和特异性的可靠诊断方法，其成本应使中低收入国家以及高收入国家能够广泛获得这些方法。基于蛋白质的诊断开发和生产依赖于作为这些测试的关键组分的关键蛋白质基试剂的可用性。这样的测试可能需要每年以数百万至数十亿的数量制造，因此需要可靠且充足的真实且低成本的蛋白质试剂供应。例如，对于简单的疟疾侧流测试，每百万次测试需要10-500 mg量的3种不同的蛋白质试剂，预计每年需要4亿次测试。这些试剂蛋白中的一些必须在培养的哺乳动物细胞系统中制备以正确折叠、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灵敏度发挥作用。尽管有一些用于从商业上可获得的哺乳动物细胞生产重组蛋白的蛋白试剂表达系统，但是这些已经被改进和开发，主要用于表达高价值蛋白质，以用作治疗一系列疾病和病症的药物。获得这样的系统涉及过高的成本，这反过来使得使用这些必需的试剂蛋白质来以必要的高容量制造诊断剂成问题。因此，在以允许其应用于商业上可行的诊断的成本获得哺乳动物细胞重组蛋白试剂方面存在大量未满足的需求。这里描述的项目将在英国领先的诊断公司Mologic内建立技术、专有技术和系统，用于在培养的哺乳动物细胞系统中快速生产足够量的重组蛋白试剂，在质量和成本上可行，应用于低成本快速诊断，在发达国家和发展中国家销售。这将确保为诊断的开发和制造提供试剂，并且还提供快速生成新的必需蛋白试剂的能力，以响应当前和新兴的需求，其成本允许其应用于诊断。

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.