biostudies-literatureCheng XHHS | National Institutes of HealthElisabeth K. Harris Foundation TrustHHS | National Institutes of Health (NIH)NIDDK NIH HHSNIDA NIH HHSHHS | NIH | National Institute of Neurological Disorders and Stroke (NINDS)HHS | NIH | National Institute of Neurological Disorders and StrokeHHS | NIH | National Institute on Drug Abuse (NIDA)HHS | NIH | National Institute on Drug AbuseNINDS NIH HHSNIGMS NIH HHSe2425526122https://www.ebi.ac.uk/biostudies/studies/S-EPMC11892595biostudies-literatureUnknown122(9)Mimicking metabolic pathways on electrodes enables in vivo metabolite monitoring for decoding metabolism. Conventional in vivo sensors cannot accommodate underlying complex reactions involving multiple enzymes and cofactors, addressing only a fraction of enzymatic reactions for few metabolites. We devised a single-wall-carbon-nanotube-electrode architecture supporting tandem metabolic pathway-like reactions linkable to oxidoreductase-based electrochemical analysis, making a vast majority of metabolites detectable in vivo. This architecture robustly integrates cofactors, self-mediates reactions at maximum enzyme capacity, and facilitates metabolite intermediation/detection and interference inactivation through multifunctional enzymatic use. Accordingly, we developed sensors targeting 12 metProceedings of the National Academy of Sciences of the United States of AmericaTandem metabolic reaction-based sensors unlock in vivo metabolomics.PMC11892595N/AR33 DK128711R35GM1382415R21DK128711-02R01DA047637R01NS115537UH3 NS119772R01 DA047637R01 NS115537R35 GM138241UH3NS119772R21 DK128711Bahramian MLei LReyes BLLin SDavis RWHojaiji HJelinek DEmaminejad SCheng XWang JHuang RPatel PGromilina EForman SMilla CZhang AYu KBPuri STan JHo CLu DCLi ZZhu JYu LWMatsumoto JHColler HALiu CVoisin LHsiao EYfalseTandem metabolic reaction-based sensors unlock in vivo metabolomics.Mimicking metabolic pathways on electrodes enables in vivo metabolite monitoring for decoding metabolism. Conventional in vivo sensors cannot accommodate underlying complex reactions involving multiple enzymes and cofactors, addressing only a fraction of enzymatic reactions for few metabolites. We devised a single-wall-carbon-nanotube-electrode architecture supporting tandem metabolic pathway-like reactions linkable to oxidoreductase-based electrochemical analysis, making a vast majority of metabolites detectable in vivo. This architecture robustly integrates cofactors, self-mediates reactions at maximum enzyme capacity, and facilitates metabolite intermediation/detection and interference inactivation through multifunctional enzymatic use. Accordingly, we developed sensors targeting 12 met2025-01-01T00:00:00Z2025 Mar2026-05-29T16:49:24.279Z2025-04-04T02:21:55.809ZS-EPMC118925954001456910.1073/pnas.2425526122