Molecularly Imprinted Nanoparticles Assay (MINA) in Pseudo ELISA: An Alternative to Detect and Quantify Octopamine in Water and Human Urine Samples

In 2004, octopamine was added to the list of drugs banned by the world anti-doping agency (WADA) and prohibited in any sport competition. This work aims to develop a new analytical method to detect octopamine in water and human urine samples. We proposed a pseudo-enzyme-linked immunosorbent assay (p...

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Veröffentlicht in:Polymers 2019-09, Vol.11 (9), p.1497
Hauptverfasser: Moczko, Ewa, Díaz, Richard, Rivas, Bernabé, García, Camilo, Pereira, Eduardo, Piletsky, Sergey, Cáceres, César
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container_end_page
container_issue 9
container_start_page 1497
container_title Polymers
container_volume 11
creator Moczko, Ewa
Díaz, Richard
Rivas, Bernabé
García, Camilo
Pereira, Eduardo
Piletsky, Sergey
Cáceres, César
description In 2004, octopamine was added to the list of drugs banned by the world anti-doping agency (WADA) and prohibited in any sport competition. This work aims to develop a new analytical method to detect octopamine in water and human urine samples. We proposed a pseudo-enzyme-linked immunosorbent assay (pseudo-ELISA) by replacing traditional monoclonal antibodies with molecularly imprinted polymer nanoparticles (nanoMIPs). NanoMIPs were synthesised by a solid-phase approach using a persulfate initiated polymerisation in water. Their performance was analysed in pseudo competitive ELISA based on the competition between free octopamine and octopamine-HRP conjugated. The final assay was able to detect octopamine in water within the range 1 nmol·L−1–0.1 mol·L−1 with a detection limit of 0.047 ± 0.00231 µg·mL−1 and in human urine samples within the range 1 nmol·L−1–0.0001 mol·L−1 with a detection limit of 0.059 ± 0.00281 µg·mL−1. In all experiments, nanoMIPs presented high affinity to the target molecules and almost no cross-reactivity with analogues of octopamine such as pseudophedrine or l-Tyrosine. Only slight interference was observed from the human urine matrix. The high affinity and specificity of nanoMIPs and no need to maintain a cold chain logistics makes the nanoMIPs a competitive alternative to antibodies. Furthermore, this work is the first attempt to use nanoMIPs in pseudo-ELISA assays to detect octopamine.
doi_str_mv 10.3390/polym11091497
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subjects Affinity
Antibodies
Antigens
Assaying
Caustic soda
Chemical synthesis
Competition
Human wastes
Imprinted polymers
Ligands
Logistics
Monoclonal antibodies
Nanoparticles
Nitrogen
Sodium
Solid phases
Tyrosine
Urine
title Molecularly Imprinted Nanoparticles Assay (MINA) in Pseudo ELISA: An Alternative to Detect and Quantify Octopamine in Water and Human Urine Samples
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