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

Molecularly Imprinted Nanoparticles Assay (MINA) in Pseudo ELISA: An Alternative to Detect and Quantify Octopamine in Water and Human Urine Samples / Ewa Moczko, Richard Díaz, Bernabé Rivas, Camilo García, Eduardo Pereira, Sergey Piletsky, César Cáceres. - (Polymers (Basel) 11 (2019) 9 (13 September); p. 1-13).
- PMID: 31540212.
- DOI: 10.3390/polym11091497


Abstract

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.

Original document

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Science
Study
Date
13 September 2019
People
Cáceres, César
Díaz, Richard
García, Camilo
Moczko, Ewa
Pereira, Eduardo
Piletsky, Sergey
Rivas, Bernabé
Country
Chile
Language
English
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Universidad Católica de la Santísima Concepción - Catholic University of the Most Holy Conception
Universidad Católica de Temuco - Temuco Catholic University
University of Leicester
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Testing method development
Doping classes
S6. Stimulants
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9 September 2019
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4 October 2019
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