An optical biosensor assay for rapid dual detection of Botulinum neurotoxins A and E
The enzymatic activity of the pathogenic botulinum neurotoxins type A and E (BoNT/A and E) leads to potentially lethal paralytic symptoms in humans and their prompt detection is of crucial importance. A chip assay based on Surface Plasmon Resonance monitoring of the cleavage products is a simple met...
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| Veröffentlicht in: | Scientific reports 2015-12, Vol.5 (1), p.17953, Article 17953 |
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| creator | Lévêque, Christian Ferracci, Géraldine Maulet, Yves Mazuet, Christelle Popoff, Michel R. Blanchard, Marie-Pierre Seagar, Michael El Far, Oussama |
| description | The enzymatic activity of the pathogenic botulinum neurotoxins type A and E (BoNT/A and E) leads to potentially lethal paralytic symptoms in humans and their prompt detection is of crucial importance. A chip assay based on Surface Plasmon Resonance monitoring of the cleavage products is a simple method that we have previously established to detect BoNT/A activity. We have now developed a similar format assay to measure BoNT/E activity. A monoclonal antibody specifically recognizing SNAP25 cleaved by BoNT/E was generated and used to measure the appearance of the neo-epitope following injection of BoNT/E over SNAP-25 immobilized on a chip. This assay detects BoNT/E activity at 1 LD
50
/ml within minutes and linear dose-responses curves were obtained using a multiplexed biosensor. A threshold of 0.01 LD
50
/ml was achieved after 5 h of cleavage. This assay is 10-fold more sensitive than the
in vivo
assay for direct detection of BoNT/E in serum samples. The SNAP25 chip assay is able to discriminate in an automated manner the presence of BoNT/E, BoNT/A or a combination of both toxins. |
| doi_str_mv | 10.1038/srep17953 |
| format | Article |
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50
/ml within minutes and linear dose-responses curves were obtained using a multiplexed biosensor. A threshold of 0.01 LD
50
/ml was achieved after 5 h of cleavage. This assay is 10-fold more sensitive than the
in vivo
assay for direct detection of BoNT/E in serum samples. The SNAP25 chip assay is able to discriminate in an automated manner the presence of BoNT/E, BoNT/A or a combination of both toxins.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/srep17953</identifier><identifier>PMID: 26648139</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>13/62 ; 631/1647/277 ; 631/378/548/2589 ; 631/61/168 ; 631/61/32 ; 631/61/350/59 ; 82/1 ; 82/103 ; 9/10 ; Antibodies, Monoclonal - immunology ; Biosensing Techniques ; Biosensors ; Biotechnology ; Botulinum Toxins - immunology ; Botulinum Toxins, Type A - immunology ; Enzymatic activity ; Enzyme Activation ; Epitopes ; Epitopes - immunology ; Human health and pathology ; Humanities and Social Sciences ; Humans ; Kinetics ; Lab-On-A-Chip Devices ; Life Sciences ; Microchip Analytical Procedures ; multidisciplinary ; Neurons and Cognition ; Neurotoxins ; Science ; Sensitivity and Specificity ; SNAP-25 protein ; Substrate Specificity ; Surface plasmon resonance ; Toxicology ; Toxins</subject><ispartof>Scientific reports, 2015-12, Vol.5 (1), p.17953, Article 17953</ispartof><rights>The Author(s) 2015</rights><rights>Copyright Nature Publishing Group Dec 2015</rights><rights>Attribution</rights><rights>Copyright © 2015, Macmillan Publishers Limited 2015 Macmillan Publishers Limited</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c475t-3229f7012091806f35bf52b7f8e868c4ab066ccf8853903d078891abdeffd0413</citedby><cites>FETCH-LOGICAL-c475t-3229f7012091806f35bf52b7f8e868c4ab066ccf8853903d078891abdeffd0413</cites><orcidid>0000-0001-7619-9050 ; 0000-0001-9305-8989</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4673697/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4673697/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,27924,27925,41120,42189,51576,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26648139$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://inserm.hal.science/inserm-01708826$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Lévêque, Christian</creatorcontrib><creatorcontrib>Ferracci, Géraldine</creatorcontrib><creatorcontrib>Maulet, Yves</creatorcontrib><creatorcontrib>Mazuet, Christelle</creatorcontrib><creatorcontrib>Popoff, Michel R.</creatorcontrib><creatorcontrib>Blanchard, Marie-Pierre</creatorcontrib><creatorcontrib>Seagar, Michael</creatorcontrib><creatorcontrib>El Far, Oussama</creatorcontrib><title>An optical biosensor assay for rapid dual detection of Botulinum neurotoxins A and E</title><title>Scientific reports</title><addtitle>Sci Rep</addtitle><addtitle>Sci Rep</addtitle><description>The enzymatic activity of the pathogenic botulinum neurotoxins type A and E (BoNT/A and E) leads to potentially lethal paralytic symptoms in humans and their prompt detection is of crucial importance. A chip assay based on Surface Plasmon Resonance monitoring of the cleavage products is a simple method that we have previously established to detect BoNT/A activity. We have now developed a similar format assay to measure BoNT/E activity. A monoclonal antibody specifically recognizing SNAP25 cleaved by BoNT/E was generated and used to measure the appearance of the neo-epitope following injection of BoNT/E over SNAP-25 immobilized on a chip. This assay detects BoNT/E activity at 1 LD
50
/ml within minutes and linear dose-responses curves were obtained using a multiplexed biosensor. A threshold of 0.01 LD
50
/ml was achieved after 5 h of cleavage. This assay is 10-fold more sensitive than the
in vivo
assay for direct detection of BoNT/E in serum samples. The SNAP25 chip assay is able to discriminate in an automated manner the presence of BoNT/E, BoNT/A or a combination of both toxins.</description><subject>13/62</subject><subject>631/1647/277</subject><subject>631/378/548/2589</subject><subject>631/61/168</subject><subject>631/61/32</subject><subject>631/61/350/59</subject><subject>82/1</subject><subject>82/103</subject><subject>9/10</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>Biosensing Techniques</subject><subject>Biosensors</subject><subject>Biotechnology</subject><subject>Botulinum Toxins - immunology</subject><subject>Botulinum Toxins, Type A - immunology</subject><subject>Enzymatic activity</subject><subject>Enzyme Activation</subject><subject>Epitopes</subject><subject>Epitopes - immunology</subject><subject>Human health and pathology</subject><subject>Humanities and Social Sciences</subject><subject>Humans</subject><subject>Kinetics</subject><subject>Lab-On-A-Chip Devices</subject><subject>Life Sciences</subject><subject>Microchip Analytical Procedures</subject><subject>multidisciplinary</subject><subject>Neurons and Cognition</subject><subject>Neurotoxins</subject><subject>Science</subject><subject>Sensitivity and Specificity</subject><subject>SNAP-25 protein</subject><subject>Substrate Specificity</subject><subject>Surface plasmon resonance</subject><subject>Toxicology</subject><subject>Toxins</subject><issn>2045-2322</issn><issn>2045-2322</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>C6C</sourceid><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNplUU1LAzEUDKKoaA_-AQl4UqzmYzcfF6GKWqHgpZ5DdjepKW2yJrui_95Ia6maSx7MvJk3DAAnGF1hRMV1iqbFXJZ0BxwSVJRDQgnZ3ZoPwCClOcqvJLLAch8cEMYKgak8BNORh6HtXK0XsHIhGZ9ChDol_QltnqJuXQObPsON6UzduZAXLLwNXb9wvl9Cb_oYuvDhfIIjqH0D74_BntWLZAbr_wi8PNxP78bDyfPj091oMqwLXnbDfJu0HGGCJBaIWVpWtiQVt8IIJupCV4ixurZClFQi2iAuhMS6aoy1DSowPQI3K922r5amqY3vol6oNrqljp8qaKd-I969qll4VwXjlEmeBS5XAq9_1sajicqBTFwqhDkSgrD3b7-ztV8Mb71JnZqHPvocUWGBUUkLxGVmna9YdQwpt2M3yhip78rUprLMPd1OsGH-FJQJFytCypCfmbhl-U_tC29Xn5k</recordid><startdate>20151209</startdate><enddate>20151209</enddate><creator>Lévêque, Christian</creator><creator>Ferracci, Géraldine</creator><creator>Maulet, Yves</creator><creator>Mazuet, Christelle</creator><creator>Popoff, Michel R.</creator><creator>Blanchard, Marie-Pierre</creator><creator>Seagar, Michael</creator><creator>El Far, Oussama</creator><general>Nature Publishing Group UK</general><general>Nature Publishing Group</general><scope>C6C</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>1XC</scope><scope>VOOES</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-7619-9050</orcidid><orcidid>https://orcid.org/0000-0001-9305-8989</orcidid></search><sort><creationdate>20151209</creationdate><title>An optical biosensor assay for rapid dual detection of Botulinum neurotoxins A and E</title><author>Lévêque, Christian ; Ferracci, Géraldine ; Maulet, Yves ; Mazuet, Christelle ; Popoff, Michel R. ; Blanchard, Marie-Pierre ; Seagar, Michael ; El Far, Oussama</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c475t-3229f7012091806f35bf52b7f8e868c4ab066ccf8853903d078891abdeffd0413</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>13/62</topic><topic>631/1647/277</topic><topic>631/378/548/2589</topic><topic>631/61/168</topic><topic>631/61/32</topic><topic>631/61/350/59</topic><topic>82/1</topic><topic>82/103</topic><topic>9/10</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>Biosensing Techniques</topic><topic>Biosensors</topic><topic>Biotechnology</topic><topic>Botulinum Toxins - immunology</topic><topic>Botulinum Toxins, Type A - immunology</topic><topic>Enzymatic activity</topic><topic>Enzyme Activation</topic><topic>Epitopes</topic><topic>Epitopes - immunology</topic><topic>Human health and pathology</topic><topic>Humanities and Social Sciences</topic><topic>Humans</topic><topic>Kinetics</topic><topic>Lab-On-A-Chip Devices</topic><topic>Life Sciences</topic><topic>Microchip Analytical Procedures</topic><topic>multidisciplinary</topic><topic>Neurons and Cognition</topic><topic>Neurotoxins</topic><topic>Science</topic><topic>Sensitivity and Specificity</topic><topic>SNAP-25 protein</topic><topic>Substrate Specificity</topic><topic>Surface plasmon resonance</topic><topic>Toxicology</topic><topic>Toxins</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lévêque, Christian</creatorcontrib><creatorcontrib>Ferracci, Géraldine</creatorcontrib><creatorcontrib>Maulet, Yves</creatorcontrib><creatorcontrib>Mazuet, Christelle</creatorcontrib><creatorcontrib>Popoff, Michel R.</creatorcontrib><creatorcontrib>Blanchard, Marie-Pierre</creatorcontrib><creatorcontrib>Seagar, Michael</creatorcontrib><creatorcontrib>El Far, Oussama</creatorcontrib><collection>Springer Nature OA Free Journals</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Biological Science Database</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 Basic</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lévêque, Christian</au><au>Ferracci, Géraldine</au><au>Maulet, Yves</au><au>Mazuet, Christelle</au><au>Popoff, Michel R.</au><au>Blanchard, Marie-Pierre</au><au>Seagar, Michael</au><au>El Far, Oussama</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>An optical biosensor assay for rapid dual detection of Botulinum neurotoxins A and E</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2015-12-09</date><risdate>2015</risdate><volume>5</volume><issue>1</issue><spage>17953</spage><pages>17953-</pages><artnum>17953</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>The enzymatic activity of the pathogenic botulinum neurotoxins type A and E (BoNT/A and E) leads to potentially lethal paralytic symptoms in humans and their prompt detection is of crucial importance. A chip assay based on Surface Plasmon Resonance monitoring of the cleavage products is a simple method that we have previously established to detect BoNT/A activity. We have now developed a similar format assay to measure BoNT/E activity. A monoclonal antibody specifically recognizing SNAP25 cleaved by BoNT/E was generated and used to measure the appearance of the neo-epitope following injection of BoNT/E over SNAP-25 immobilized on a chip. This assay detects BoNT/E activity at 1 LD
50
/ml within minutes and linear dose-responses curves were obtained using a multiplexed biosensor. A threshold of 0.01 LD
50
/ml was achieved after 5 h of cleavage. This assay is 10-fold more sensitive than the
in vivo
assay for direct detection of BoNT/E in serum samples. The SNAP25 chip assay is able to discriminate in an automated manner the presence of BoNT/E, BoNT/A or a combination of both toxins.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>26648139</pmid><doi>10.1038/srep17953</doi><orcidid>https://orcid.org/0000-0001-7619-9050</orcidid><orcidid>https://orcid.org/0000-0001-9305-8989</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | 13/62 631/1647/277 631/378/548/2589 631/61/168 631/61/32 631/61/350/59 82/1 82/103 9/10 Antibodies, Monoclonal - immunology Biosensing Techniques Biosensors Biotechnology Botulinum Toxins - immunology Botulinum Toxins, Type A - immunology Enzymatic activity Enzyme Activation Epitopes Epitopes - immunology Human health and pathology Humanities and Social Sciences Humans Kinetics Lab-On-A-Chip Devices Life Sciences Microchip Analytical Procedures multidisciplinary Neurons and Cognition Neurotoxins Science Sensitivity and Specificity SNAP-25 protein Substrate Specificity Surface plasmon resonance Toxicology Toxins |
| title | An optical biosensor assay for rapid dual detection of Botulinum neurotoxins A and E |
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