Enzyme inhibition-based biosensor for the electrochemical detection of microcystins in natural blooms of cyanobacteria
An electrochemical biosensor for the detection of microcystin has been developed based on the inhibition of the protein phosphatase 2A (PP2A) by this cyanobacterial toxin. The enzyme has been immobilised by entrapment using a poly(vinyl alcohol) azide-unit pendant water-soluble photopolymer (PVA-AWP...
Gespeichert in:
| Veröffentlicht in: | Talanta (Oxford) 2007-04, Vol.72 (1), p.179-186 |
|---|---|
| Hauptverfasser: | , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 186 |
|---|---|
| container_issue | 1 |
| container_start_page | 179 |
| container_title | Talanta (Oxford) |
| container_volume | 72 |
| creator | Campàs, Mònica Szydłowska, Dorota Trojanowicz, Marek Marty, Jean-Louis |
| description | An electrochemical biosensor for the detection of microcystin has been developed based on the inhibition of the protein phosphatase 2A (PP2A) by this cyanobacterial toxin. The enzyme has been immobilised by entrapment using a poly(vinyl alcohol) azide-unit pendant water-soluble photopolymer (PVA-AWP). Electrode supports and immobilisation conditions have been optimised by colorimetric assays, the highest immobilisation yields being obtained with screen-printed graphite electrodes and the 1:2 PP2A:PVA ratio. Catechyl monophosphate (CMP), α-naphthyl phosphate (α-NP) and 4-methylumbelliferyl phosphate (4-MUP) have been used as phosphorylated substrates to monitor the protein phosphatase activity by electrochemical methods, the former providing the highest chronoamperometric currents at appropriate working potentials (+450
mV
versus Ag/AgCl). Incubation with standard microcystin solutions has demonstrated the inhibition of the immobilised enzyme, proportional to the toxin concentration. The standard inhibition curve has provided a 50% inhibition coefficient (IC
50) of 83
μg
L
−1, a limit of detection (LOD; 35% inhibition) of 37
μg
L
−1, and 100% inhibition at about 1000
μg
L
−1. Real samples of cyanobacterial blooms from the Tarn River (Midi-Pyrénées, France) have been analysed using the developed amperometric biosensor and the toxin contents have been compared to those obtained by a conventional colorimetric protein phosphatase inhibition (PPI) assay and high-performance liquid chromatography (HPLC). The results clearly justify the use of the developed amperometric biosensor as screening method for microcystin detection. |
| doi_str_mv | 10.1016/j.talanta.2006.10.012 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_733878020</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S0039914006006783</els_id><sourcerecordid>20040384</sourcerecordid><originalsourceid>FETCH-LOGICAL-c491t-eb513d913bcf7006fc0be23c9f7f91b0de769e0038e6df1e13075d4be6e9f773</originalsourceid><addsrcrecordid>eNqFkU9r3DAQxUVoSLZpP0KLL21P3o4s27JOJYT0DwR6yd1I8ojVYkuppA1sP31mWUNu7UEIRr83o3mPsQ8cthx4_3W_LXrWoehtA9BTbQu8uWAbPkhRi06KN2wDIFSteAvX7G3OewBoBIgrds0VSN4ptWHP9-HvccHKh503vvgYaqMzTpXxMWPIMVWOTtlhhTPakqLd4eKtnqsJCxVIUUVXUYmejrn4kKlZFXQ5JILMHOOST4Q96hCNtgWT1-_YpdNzxvfrfcMev98_3v2sH37_-HV3-1DbVvFSo-m4mBQXxjpJazoLBhthlZNOcQMTyl4hrTlgPzmOXIDsptZgj4RIccO-nNs-pfjngLmMi88WZzIO4yGPUohBDtAAkZ__SZLJLY1pCezOIO2bc0I3PiW_6HQcOYynZMb9uCZzEvWnMiVDuo_rgINZcHpVrVEQ8GkFdCZ7XdLB-vzKDX3XqqYj7tuZQ_Lt2WMas_UYLE4-UR7jFP1_vvIC6yqxtA</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>20040384</pqid></control><display><type>article</type><title>Enzyme inhibition-based biosensor for the electrochemical detection of microcystins in natural blooms of cyanobacteria</title><source>Access via ScienceDirect (Elsevier)</source><creator>Campàs, Mònica ; Szydłowska, Dorota ; Trojanowicz, Marek ; Marty, Jean-Louis</creator><creatorcontrib>Campàs, Mònica ; Szydłowska, Dorota ; Trojanowicz, Marek ; Marty, Jean-Louis</creatorcontrib><description>An electrochemical biosensor for the detection of microcystin has been developed based on the inhibition of the protein phosphatase 2A (PP2A) by this cyanobacterial toxin. The enzyme has been immobilised by entrapment using a poly(vinyl alcohol) azide-unit pendant water-soluble photopolymer (PVA-AWP). Electrode supports and immobilisation conditions have been optimised by colorimetric assays, the highest immobilisation yields being obtained with screen-printed graphite electrodes and the 1:2 PP2A:PVA ratio. Catechyl monophosphate (CMP), α-naphthyl phosphate (α-NP) and 4-methylumbelliferyl phosphate (4-MUP) have been used as phosphorylated substrates to monitor the protein phosphatase activity by electrochemical methods, the former providing the highest chronoamperometric currents at appropriate working potentials (+450
mV
versus Ag/AgCl). Incubation with standard microcystin solutions has demonstrated the inhibition of the immobilised enzyme, proportional to the toxin concentration. The standard inhibition curve has provided a 50% inhibition coefficient (IC
50) of 83
μg
L
−1, a limit of detection (LOD; 35% inhibition) of 37
μg
L
−1, and 100% inhibition at about 1000
μg
L
−1. Real samples of cyanobacterial blooms from the Tarn River (Midi-Pyrénées, France) have been analysed using the developed amperometric biosensor and the toxin contents have been compared to those obtained by a conventional colorimetric protein phosphatase inhibition (PPI) assay and high-performance liquid chromatography (HPLC). The results clearly justify the use of the developed amperometric biosensor as screening method for microcystin detection.</description><identifier>ISSN: 0039-9140</identifier><identifier>EISSN: 1873-3573</identifier><identifier>DOI: 10.1016/j.talanta.2006.10.012</identifier><identifier>PMID: 19071599</identifier><identifier>CODEN: TLNTA2</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Analytical chemistry ; Biological and medical sciences ; Biosensors ; Biotechnology ; Catechyl monophosphate (CMP) ; Chemistry ; Chromatographic methods and physical methods associated with chromatography ; Cyanobacteria ; Electrochemical detection ; Electrochemical methods ; Exact sciences and technology ; Fundamental and applied biological sciences. Psychology ; Inhibition ; Methods. Procedures. Technologies ; Microcystin (MC) ; Other chromatographic methods ; Poly(vinyl alcohol) azide-unit pendant water-soluble photopolymer (PVA-AWP) Entrapment ; Protein phosphatase 2A (PP2A) ; Spectrometric and optical methods ; Various methods and equipments</subject><ispartof>Talanta (Oxford), 2007-04, Vol.72 (1), p.179-186</ispartof><rights>2006 Elsevier B.V.</rights><rights>2007 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c491t-eb513d913bcf7006fc0be23c9f7f91b0de769e0038e6df1e13075d4be6e9f773</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.talanta.2006.10.012$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18654925$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19071599$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Campàs, Mònica</creatorcontrib><creatorcontrib>Szydłowska, Dorota</creatorcontrib><creatorcontrib>Trojanowicz, Marek</creatorcontrib><creatorcontrib>Marty, Jean-Louis</creatorcontrib><title>Enzyme inhibition-based biosensor for the electrochemical detection of microcystins in natural blooms of cyanobacteria</title><title>Talanta (Oxford)</title><addtitle>Talanta</addtitle><description>An electrochemical biosensor for the detection of microcystin has been developed based on the inhibition of the protein phosphatase 2A (PP2A) by this cyanobacterial toxin. The enzyme has been immobilised by entrapment using a poly(vinyl alcohol) azide-unit pendant water-soluble photopolymer (PVA-AWP). Electrode supports and immobilisation conditions have been optimised by colorimetric assays, the highest immobilisation yields being obtained with screen-printed graphite electrodes and the 1:2 PP2A:PVA ratio. Catechyl monophosphate (CMP), α-naphthyl phosphate (α-NP) and 4-methylumbelliferyl phosphate (4-MUP) have been used as phosphorylated substrates to monitor the protein phosphatase activity by electrochemical methods, the former providing the highest chronoamperometric currents at appropriate working potentials (+450
mV
versus Ag/AgCl). Incubation with standard microcystin solutions has demonstrated the inhibition of the immobilised enzyme, proportional to the toxin concentration. The standard inhibition curve has provided a 50% inhibition coefficient (IC
50) of 83
μg
L
−1, a limit of detection (LOD; 35% inhibition) of 37
μg
L
−1, and 100% inhibition at about 1000
μg
L
−1. Real samples of cyanobacterial blooms from the Tarn River (Midi-Pyrénées, France) have been analysed using the developed amperometric biosensor and the toxin contents have been compared to those obtained by a conventional colorimetric protein phosphatase inhibition (PPI) assay and high-performance liquid chromatography (HPLC). The results clearly justify the use of the developed amperometric biosensor as screening method for microcystin detection.</description><subject>Analytical chemistry</subject><subject>Biological and medical sciences</subject><subject>Biosensors</subject><subject>Biotechnology</subject><subject>Catechyl monophosphate (CMP)</subject><subject>Chemistry</subject><subject>Chromatographic methods and physical methods associated with chromatography</subject><subject>Cyanobacteria</subject><subject>Electrochemical detection</subject><subject>Electrochemical methods</subject><subject>Exact sciences and technology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Inhibition</subject><subject>Methods. Procedures. Technologies</subject><subject>Microcystin (MC)</subject><subject>Other chromatographic methods</subject><subject>Poly(vinyl alcohol) azide-unit pendant water-soluble photopolymer (PVA-AWP) Entrapment</subject><subject>Protein phosphatase 2A (PP2A)</subject><subject>Spectrometric and optical methods</subject><subject>Various methods and equipments</subject><issn>0039-9140</issn><issn>1873-3573</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><recordid>eNqFkU9r3DAQxUVoSLZpP0KLL21P3o4s27JOJYT0DwR6yd1I8ojVYkuppA1sP31mWUNu7UEIRr83o3mPsQ8cthx4_3W_LXrWoehtA9BTbQu8uWAbPkhRi06KN2wDIFSteAvX7G3OewBoBIgrds0VSN4ptWHP9-HvccHKh503vvgYaqMzTpXxMWPIMVWOTtlhhTPakqLd4eKtnqsJCxVIUUVXUYmejrn4kKlZFXQ5JILMHOOST4Q96hCNtgWT1-_YpdNzxvfrfcMev98_3v2sH37_-HV3-1DbVvFSo-m4mBQXxjpJazoLBhthlZNOcQMTyl4hrTlgPzmOXIDsptZgj4RIccO-nNs-pfjngLmMi88WZzIO4yGPUohBDtAAkZ__SZLJLY1pCezOIO2bc0I3PiW_6HQcOYynZMb9uCZzEvWnMiVDuo_rgINZcHpVrVEQ8GkFdCZ7XdLB-vzKDX3XqqYj7tuZQ_Lt2WMas_UYLE4-UR7jFP1_vvIC6yqxtA</recordid><startdate>20070415</startdate><enddate>20070415</enddate><creator>Campàs, Mònica</creator><creator>Szydłowska, Dorota</creator><creator>Trojanowicz, Marek</creator><creator>Marty, Jean-Louis</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QH</scope><scope>7QO</scope><scope>7U7</scope><scope>7UA</scope><scope>8FD</scope><scope>C1K</scope><scope>F1W</scope><scope>FR3</scope><scope>H97</scope><scope>H99</scope><scope>L.F</scope><scope>L.G</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20070415</creationdate><title>Enzyme inhibition-based biosensor for the electrochemical detection of microcystins in natural blooms of cyanobacteria</title><author>Campàs, Mònica ; Szydłowska, Dorota ; Trojanowicz, Marek ; Marty, Jean-Louis</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c491t-eb513d913bcf7006fc0be23c9f7f91b0de769e0038e6df1e13075d4be6e9f773</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Analytical chemistry</topic><topic>Biological and medical sciences</topic><topic>Biosensors</topic><topic>Biotechnology</topic><topic>Catechyl monophosphate (CMP)</topic><topic>Chemistry</topic><topic>Chromatographic methods and physical methods associated with chromatography</topic><topic>Cyanobacteria</topic><topic>Electrochemical detection</topic><topic>Electrochemical methods</topic><topic>Exact sciences and technology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Inhibition</topic><topic>Methods. Procedures. Technologies</topic><topic>Microcystin (MC)</topic><topic>Other chromatographic methods</topic><topic>Poly(vinyl alcohol) azide-unit pendant water-soluble photopolymer (PVA-AWP) Entrapment</topic><topic>Protein phosphatase 2A (PP2A)</topic><topic>Spectrometric and optical methods</topic><topic>Various methods and equipments</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Campàs, Mònica</creatorcontrib><creatorcontrib>Szydłowska, Dorota</creatorcontrib><creatorcontrib>Trojanowicz, Marek</creatorcontrib><creatorcontrib>Marty, Jean-Louis</creatorcontrib><collection>Pascal-Francis</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aqualine</collection><collection>Biotechnology Research Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Water Resources Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Engineering Research Database</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>ASFA: Marine Biotechnology Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Marine Biotechnology Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Talanta (Oxford)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Campàs, Mònica</au><au>Szydłowska, Dorota</au><au>Trojanowicz, Marek</au><au>Marty, Jean-Louis</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enzyme inhibition-based biosensor for the electrochemical detection of microcystins in natural blooms of cyanobacteria</atitle><jtitle>Talanta (Oxford)</jtitle><addtitle>Talanta</addtitle><date>2007-04-15</date><risdate>2007</risdate><volume>72</volume><issue>1</issue><spage>179</spage><epage>186</epage><pages>179-186</pages><issn>0039-9140</issn><eissn>1873-3573</eissn><coden>TLNTA2</coden><abstract>An electrochemical biosensor for the detection of microcystin has been developed based on the inhibition of the protein phosphatase 2A (PP2A) by this cyanobacterial toxin. The enzyme has been immobilised by entrapment using a poly(vinyl alcohol) azide-unit pendant water-soluble photopolymer (PVA-AWP). Electrode supports and immobilisation conditions have been optimised by colorimetric assays, the highest immobilisation yields being obtained with screen-printed graphite electrodes and the 1:2 PP2A:PVA ratio. Catechyl monophosphate (CMP), α-naphthyl phosphate (α-NP) and 4-methylumbelliferyl phosphate (4-MUP) have been used as phosphorylated substrates to monitor the protein phosphatase activity by electrochemical methods, the former providing the highest chronoamperometric currents at appropriate working potentials (+450
mV
versus Ag/AgCl). Incubation with standard microcystin solutions has demonstrated the inhibition of the immobilised enzyme, proportional to the toxin concentration. The standard inhibition curve has provided a 50% inhibition coefficient (IC
50) of 83
μg
L
−1, a limit of detection (LOD; 35% inhibition) of 37
μg
L
−1, and 100% inhibition at about 1000
μg
L
−1. Real samples of cyanobacterial blooms from the Tarn River (Midi-Pyrénées, France) have been analysed using the developed amperometric biosensor and the toxin contents have been compared to those obtained by a conventional colorimetric protein phosphatase inhibition (PPI) assay and high-performance liquid chromatography (HPLC). The results clearly justify the use of the developed amperometric biosensor as screening method for microcystin detection.</abstract><cop>Amsterdam</cop><cop>Oxford</cop><pub>Elsevier B.V</pub><pmid>19071599</pmid><doi>10.1016/j.talanta.2006.10.012</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0039-9140 |
| ispartof | Talanta (Oxford), 2007-04, Vol.72 (1), p.179-186 |
| issn | 0039-9140 1873-3573 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_733878020 |
| source | Access via ScienceDirect (Elsevier) |
| subjects | Analytical chemistry Biological and medical sciences Biosensors Biotechnology Catechyl monophosphate (CMP) Chemistry Chromatographic methods and physical methods associated with chromatography Cyanobacteria Electrochemical detection Electrochemical methods Exact sciences and technology Fundamental and applied biological sciences. Psychology Inhibition Methods. Procedures. Technologies Microcystin (MC) Other chromatographic methods Poly(vinyl alcohol) azide-unit pendant water-soluble photopolymer (PVA-AWP) Entrapment Protein phosphatase 2A (PP2A) Spectrometric and optical methods Various methods and equipments |
| title | Enzyme inhibition-based biosensor for the electrochemical detection of microcystins in natural blooms of cyanobacteria |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-29T21%3A19%3A25IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Enzyme%20inhibition-based%20biosensor%20for%20the%20electrochemical%20detection%20of%20microcystins%20in%20natural%20blooms%20of%20cyanobacteria&rft.jtitle=Talanta%20(Oxford)&rft.au=Camp%C3%A0s,%20M%C3%B2nica&rft.date=2007-04-15&rft.volume=72&rft.issue=1&rft.spage=179&rft.epage=186&rft.pages=179-186&rft.issn=0039-9140&rft.eissn=1873-3573&rft.coden=TLNTA2&rft_id=info:doi/10.1016/j.talanta.2006.10.012&rft_dat=%3Cproquest_cross%3E20040384%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=20040384&rft_id=info:pmid/19071599&rft_els_id=S0039914006006783&rfr_iscdi=true |