Automated Electrical Quantification of Vitamin B1 in a Bodily Fluid using an Engineered Nanopore-Sensor

Florian Leonardus Rudolfus Lucas, Tjemme Rinze Cornelis Piso, Nieck Jordy van der Heide, Nicole Stéphanie Galenkamp, Jos Hermans, Carsten Wloka*, Giovanni Maglia*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

11 Citations (Scopus)
25 Downloads (Pure)

Abstract

The ability to measure the concentration of metabolites in biological samples is important, both in the clinic and for home diagnostics. Here we present a nanopore-based biosensor and automated data analysis for quantification of thiamine in urine in less than a minute, without the need for recalibration. For this we use the Cytolysin A nanopore and equip it with an engineered periplasmic thiamine binding protein (TbpA). To allow fast measurements we tuned the affinity of TbpA for thiamine by redesigning the π-π stacking interactions between the thiazole group of thiamine and TbpA. This substitution resulted furthermore in a marked difference between unbound and bound state, allowing the reliable discrimination of thiamine from its two phosphorylated forms by residual current only. Using an array of nanopores, this allows the quantification within seconds, paving the way for next-generation single-molecule metabolite detection systems.

Original languageEnglish
Pages (from-to) 22849-22855
Number of pages6
JournalAngewandte Chemie (International ed. in English)
Volume60
Issue number42
Early online date14-Aug-2021
DOIs
Publication statusPublished - 11-Oct-2021

Cite this