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

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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.