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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Authors
GARCIA SANHUEZA, CAMILO ANDRES
Moczko, Ewa
Díaz, Richard
Rivas, Bernabé Luis Q.
García, Camilo
Pereira, Eduardo D.
Piletsky, Sergey A.
Cáceres, César
Moczko, Ewa
Díaz, Richard
Rivas, Bernabé Luis Q.
García, Camilo
Pereira, Eduardo D.
Piletsky, Sergey A.
Cáceres, César
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Date
Datos de publicación:
10.3390/polym11091497
Keywords
Doping - Elisa - Molecularly Imprinted Nanoparticles Assay - Molecularly Imprinted Polymers - Octopamine - Amino Acids - Doping (additives) - Molecules - Monoclonal Antibodies - Nanoparticles - Cold Chain Logistics - Elisa - Enzyme Linked Immunosorbent Assay - Human Urine Samples - Molecularly Imprinted - Molecularly Imprinted Polymer - Octopamine - World Anti-doping Agency - Drug Delivery
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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 ?g·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 ?g·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. © 2019 Elsevier B.V., All rights reserved.
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Keywords
Doping , Elisa , Molecularly Imprinted Nanoparticles Assay , Molecularly Imprinted Polymers , Octopamine , Amino Acids , Doping (additives) , Molecules , Monoclonal Antibodies , Nanoparticles , Cold Chain Logistics , Elisa , Enzyme Linked Immunosorbent Assay , Human Urine Samples , Molecularly Imprinted , Molecularly Imprinted Polymer , Octopamine , World Anti-doping Agency , Drug Delivery
Citation
10.3390/polym11091497