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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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. |
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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.</description><identifier>ISSN: 2073-4360</identifier><identifier>EISSN: 2073-4360</identifier><identifier>DOI: 10.3390/polym11091497</identifier><identifier>PMID: 31540212</identifier><language>eng</language><publisher>Basel: MDPI AG</publisher><subject>Affinity ; Antibodies ; Antigens ; Assaying ; Caustic soda ; Chemical synthesis ; Competition ; Human wastes ; Imprinted polymers ; Ligands ; Logistics ; Monoclonal antibodies ; Nanoparticles ; Nitrogen ; Sodium ; Solid phases ; Tyrosine ; Urine</subject><ispartof>Polymers, 2019-09, Vol.11 (9), p.1497</ispartof><rights>2019 by the authors. Licensee MDPI, Basel, Switzerland. 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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.</description><subject>Affinity</subject><subject>Antibodies</subject><subject>Antigens</subject><subject>Assaying</subject><subject>Caustic soda</subject><subject>Chemical synthesis</subject><subject>Competition</subject><subject>Human wastes</subject><subject>Imprinted polymers</subject><subject>Ligands</subject><subject>Logistics</subject><subject>Monoclonal antibodies</subject><subject>Nanoparticles</subject><subject>Nitrogen</subject><subject>Sodium</subject><subject>Solid phases</subject><subject>Tyrosine</subject><subject>Urine</subject><issn>2073-4360</issn><issn>2073-4360</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><recordid>eNpdkc1u1TAQhS0EolXpkr0lNmUR8E9sJyyQolLaK922oFKxjIw9gVSOndpOpTwHL1xfWiGKN2Npvjkzcwah15S847wl7-fg1olS0tK6Vc_QPiOKVzWX5Pk__z10mNINKa8WUlL1Eu1xKmrCKNtHv8-DA7M4Hd2KN9McR5_B4gvtw6xjHo2DhLuU9IqPzjcX3Vs8evwlwWIDPtlurroPuPO4cxmi13m8A5wD_gQZTMbaW_x10T6Pw4ovTS6K0-hhp_Bdl4I_wNkyaY-v4y5xpae59HuFXgzaJTh8jAfo-vPJt-Ozant5ujnutpXhLcuVNGUfzhkBO2gBbc2pbFQztFw1Qg1SUQuiNpYTzpvGakGJZMUQaCRpRWv5Afr4oDsvPyawBnyO2vXFg0nHtQ967J9m_Pir_xnueqkaUtoVgaNHgRhuF0i5n8ZkwDntISypZ6wVdVPXYoe--Q-9CUuxzBVKCMVYWYUWqnqgTAwpRRj-DkNJv7t4_-Ti_B5tGp0D</recordid><startdate>20190913</startdate><enddate>20190913</enddate><creator>Moczko, Ewa</creator><creator>Díaz, Richard</creator><creator>Rivas, Bernabé</creator><creator>García, Camilo</creator><creator>Pereira, Eduardo</creator><creator>Piletsky, Sergey</creator><creator>Cáceres, César</creator><general>MDPI AG</general><general>MDPI</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7SR</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>HCIFZ</scope><scope>JG9</scope><scope>KB.</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-2324-6735</orcidid><orcidid>https://orcid.org/0000-0002-6406-0796</orcidid></search><sort><creationdate>20190913</creationdate><title>Molecularly Imprinted Nanoparticles Assay (MINA) in Pseudo ELISA: An Alternative to Detect and Quantify Octopamine in Water and Human Urine Samples</title><author>Moczko, Ewa ; Díaz, Richard ; Rivas, Bernabé ; García, Camilo ; Pereira, Eduardo ; Piletsky, Sergey ; Cáceres, César</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c392t-6c4563320edfa5e94316878f937857f671de54cd303388da51062436e860959d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Affinity</topic><topic>Antibodies</topic><topic>Antigens</topic><topic>Assaying</topic><topic>Caustic soda</topic><topic>Chemical synthesis</topic><topic>Competition</topic><topic>Human wastes</topic><topic>Imprinted polymers</topic><topic>Ligands</topic><topic>Logistics</topic><topic>Monoclonal antibodies</topic><topic>Nanoparticles</topic><topic>Nitrogen</topic><topic>Sodium</topic><topic>Solid phases</topic><topic>Tyrosine</topic><topic>Urine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Moczko, Ewa</creatorcontrib><creatorcontrib>Díaz, Richard</creatorcontrib><creatorcontrib>Rivas, Bernabé</creatorcontrib><creatorcontrib>García, Camilo</creatorcontrib><creatorcontrib>Pereira, Eduardo</creatorcontrib><creatorcontrib>Piletsky, Sergey</creatorcontrib><creatorcontrib>Cáceres, César</creatorcontrib><collection>CrossRef</collection><collection>Engineered Materials Abstracts</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>SciTech Premium Collection</collection><collection>Materials Research Database</collection><collection>Materials Science Database</collection><collection>Materials Science Collection</collection><collection>Access via ProQuest (Open Access)</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Polymers</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Moczko, Ewa</au><au>Díaz, Richard</au><au>Rivas, Bernabé</au><au>García, Camilo</au><au>Pereira, Eduardo</au><au>Piletsky, Sergey</au><au>Cáceres, César</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecularly Imprinted Nanoparticles Assay (MINA) in Pseudo ELISA: An Alternative to Detect and Quantify Octopamine in Water and Human Urine Samples</atitle><jtitle>Polymers</jtitle><date>2019-09-13</date><risdate>2019</risdate><volume>11</volume><issue>9</issue><spage>1497</spage><pages>1497-</pages><issn>2073-4360</issn><eissn>2073-4360</eissn><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. 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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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