MicroTotal Pre Analytical Systems (MTPAS): Near-patient Approach to the Preparation of Circulating Biomarkers for Next-Generation Sensing

由于寿命延长和人口老龄化，对血液取样要求的需求增加。循环无细胞核酸（cfNAs）作为癌症、无创产前检测、器官移植监测等生物标志物的使用越来越受欢迎，因为它是非侵入性的（简单的抽血），容易重复，可以快速建立诊断、预后，甚至用于治疗应用。例如，所谓的“液体活检”和cfDNA分析可以让全球每年诊断出的1400万癌症患者获得更早的诊断和优化的治疗。然而，尽管有许多转化研究项目，cfNA的检测目前尚未在日常临床实践中实施。这其中出现了几个原因，包括(i)难以控制从血液采样到生物标志物分析的不同生物、环境和物流参数（ii）当前可用技术的成本限制了可及性（iii）对患者和临床医生有用所需的周转时间。目前的样品制备解决方案是多步骤的，可以引入变化和导致错误的诊断。此外，这些解决方案耗时，不适合在抽血后近距离提取患者，并且需要训练有素的技术人员。提取无细胞循环标记物的优化是将其从研究环境转化为临床部署的关键。缺乏工程解决方案来解决循环无细胞核酸提取的特殊性，这是该计划的基础。为了满足未来医疗保健行业的需求，提出的工作将集成新型先进材料，如电纺丝纤维，与芯片上的试剂包装在微流控盒中，从血液中提取cfna。在接近病人的情况下，这项技术将保护生物标志物免受酶的降解，并使它们与样本中存在的其他核酸相结合，从而实现无与伦比的标准化和即时保存真实的疾病状态，直到分析。拟议的解决方案涉及使用一次性药筒，如果实施，可能会产生大量额外的医疗废物。因此，该计划具有独特的可持续制造组件，研究使用天然衍生塑料（聚乳酸）来制作原型和制造低碳足迹，一次性微流体盒，可能适用于大范围的护理点设备。该项目开发的解决方案有可能显著降低循环生物标志物领域样品制备的总体成本，并以可持续的方式增加一系列生物标志物在临床诊断、生物标志物和药物发现方面的直接应用的稳健性和可靠性。与新颖的传感解决方案相结合，这项工作有可能通过优化治疗和更好的护理管理，从更早、更快的诊断中提高生活质量。到2020年，全球液体活检市场预计将达到45亿美元，前端样品制备是英国经济的一个重要领域。

Increased demand on blood sampling requirements has arisen from prolong lifespan and ageing populations. The use of circulating cell-free nucleic acids (cfNAs) as biomarkers for cancer, non-invasive prenatal testing, organ transplant monitoring and more, has grown in popularity since it is non-invasive (simple blood draw) and easily repeated, allowing the possibility to rapidly establish a diagnosis, a prognosis and even used for theranostic applications. So called "liquid biopsies" and cfDNA analysis could for example allow the fourteen millions of cancer patients diagnosed globally each year to access earlier diagnosis and optimised treatments.However, despite numerous translational research programs the detection of cfNA is not currently implemented clinically in daily practice. Several reasons have been emerged for this, including (i) the difficulty in controlling the different biological, environmental and logistic parameters from blood sampling to the analysis of biomarkers (ii) the cost of the currently available techniques, which limit accessibility (iii) the turn-around time needed to be useful for patients and clinicians. Current sample preparation solutions are multi-step which can introduce variations and lead to an erroneous diagnosis. Additionally these solutions are time consuming, not amenable to near-patient extraction following blood draw, and require highly trained technicians. The optimisation of the extraction of cell-free circulating markers is key to their translation from the research setting to clinical deployment. The lack of engineering solutions to address the specificities of circulating cell-free nucleic acid extraction, underpins this programme. To meet the requirements of future healthcare industry, the work proposed will integrate novel advanced materials such as electrospun fibres, packaged with on-chip reagents in a microfluidic cartridge to extract cfNAs from blood. Deployed near-patient, this technology will protect the biomarkers from enzymatic degradation and enrich them against the rest of the nucleic acids present in the sample, allowing an unparalleled standardisation and instant preservation of the true disease state until analysis. The solution proposed involves the use of single-use cartridges, and could generate a significant amount of additional medical waste if implemented. Therefore this programme has a unique sustainable manufacturing component, looking into the use of a naturally-derived plastic (poly-lactic acid) to prototype and manufacture low carbon footprint, disposable, microfluidic cartridges, potential applicable to a large range of point-of-care devices. The solutions developed in this programme have the potential to significantly reduce the overall cost of sample preparation in the field of circulating biomarkers, as well as increasing the robustness and reliability of a range of biomarkers with direct application in clinical diagnostic and biomarker and drug discovery, in a sustainable fashion. Hand-in-hand with novel sensing solutions, this work has the potential to increase life quality from earlier, quicker diagnosis through optimised treatment and better care management. With the global liquid biopsy market forecasts to reach $4.5 billion by 2020, front-end sample preparation constitutes an important area for the UK economy.