Development of a novel vitamin B12 point-of-care test enabled by cell-free produced functional enzymes

维生素B12缺乏症在英国乃至全球都是一个严重的健康问题，缺乏维生素B12对婴儿、儿童和成人的健康极为有害。维生素B12对大脑发育很重要，严重缺乏会导致癫痫发作和永久性脑损伤。婴儿和老人的风险很大。目前，英国每年有66万新生儿出生；其中25%是纯母乳喂养的，有B12缺乏的风险。维生素B12缺乏症的患病率随着年龄的增长而增加，在英国，65岁以上的老年人中约有20%缺乏维生素B12。对即时B12缺乏测试的需求越来越大。在这个项目中，我们将利用工程生物学技术开发一种新的B12即时检测方法。具体来说，在这个项目中，我们将开发一种新的横向流动装置，能够通过测量纸系统上的反应活性来识别非蛋白基靶标。商业侧流测试主要使用抗体来检测分析物，而不能检测抗体不能产生的非蛋白质靶标。通常，B12缺乏症的检测需要一个中心实验室和昂贵的设备。该项目将采用合成生物学无细胞系统和基因工程来生产一种功能性酶，以便在基于纸张的系统中检测B12缺乏症标记物。

Vitamin B12 deficiency is a significant health concern in the UK, and globally, with deficiency being extremely detrimental to infant, child and adult health. Vitamin B12 is important for brain development, with severe deficiency leading to seizures and permanent brain damage. Infants and the elderly are of significant risk. There are currently \>660,000 newborns born every year in the UK; 25% of which are exclusively breastfed and are at-risk of B12 deficiency. Vitamin B12 deficiency prevalence increases with age with approximately 20% of people older than 65 being deficient in the UK. There is a growing need for a point-of-care B12 deficiency test.In this project we will develop a novel B12 point-of-care test by exploiting engineering biology techniques. Specifically, in this project we will develop a novel lateral flow device capable of identifying non-protein based targets through measurement of reaction activity on a paper system. Commercial lateral flow tests principally use antibodies to detect analytes, and cannot detect non-protein targets that an antibody cannot be generated against. Typically detection of B12 deficiency requires a central lab and expensive equipment. This project will employ the synthetic biology cell-free system and genetic engineering to produce a functional enzyme to enable detection of a B12 deficiency marker in a paper based system.