Engineered nanopore biosensor for thiamine quantification: Data analysis

Biological nanopores equipped with substrate-binding proteins are powerful biosensor pairs capable of measuring low-abundance molecules in complex samples. One example of such a system pairs the thiamine-binding protein (TbpA) and the Cytolysin A (ClyA) nanopore. TbpA has been evolved to selectively bind thiamine in even the most complex samples. When a ClyA nanopore traps TbpA, even a single experiment can generate a vast amount of data in a very short time. While at the research level, even a single nanopore can accurately quantify thiamine concentrations, the setup time prohibits a practical application, such as in point-of-care devices. Such devices would likely house hundreds to millions of pores, rendering manual data analysis impossible. To bridge the gap between biosensor development and practical devices, it is therefore necessary to develop algorithms which can analyze and interpret data automatically. Here we outline our approach to nanopore data analysis to tackle the challenges in automatic analysis.