1. Automated Electrical Quantification of Vitamin B1 in a Bodily Fluid using an Engineered Nanopore Sensor
- Author
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Nicole Stéphanie Galenkamp, Carsten Wloka, Giovanni Maglia, Tjemme Rinze Cornelis Piso, Jos Hermans, Nieck Jordy van der Heide, Florian Leonardus Rudolfus Lucas, Chemical Biology 1, and Analytical Biochemistry
- Subjects
Vitamin ,Metabolite ,nanopores ,Stacking ,Biosensing Techniques ,Catalysis ,thiamine ,chemistry.chemical_compound ,Electricity ,Humans ,Thiazole ,Research Articles ,nanotechnology ,Perforin ,food and beverages ,General Medicine ,Biosensors | Hot Paper ,General Chemistry ,Periplasmic space ,biosensors ,Body Fluids ,Nanopore ,chemistry ,bayesian ,Biophysics ,Thiamine ,Carrier Proteins ,human activities ,Biosensor ,Protein Binding ,Research Article - 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 will allow the quantification within seconds, paving the way for next‐generation single‐molecule metabolite detection systems., Quantification of small molecules in biological samples is challenging. Using the concentration‐dependent current fluctuations elicited by a thiamine binding protein inside a ClyA nanopore, we show the automated quantification of thiamine in urine in less than a minute. By swapping the internal biosensor, this principle can be applied for the development of multiple next‐generation home‐diagnostic small‐molecule sensors.
- Published
- 2021