Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection

Autores
Alba Posse, Jorge Ezequiel; Gonzalez, Carolina; Carriquiriborde, Pedro; Nadra, Alejandro Daniel; Gasulla, Javier
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The presence of cyanobacterial toxins in freshwater constitutes an increasing public healthconcern, especially affecting developing countries where the high cost of available methodsmakes monitoring programs difficult. The phosphatase inhibition assay (PPIAs) is a sensitivemethod with low instrument requirements that allows the quantification of the most frequentcyanotoxins, microcystins (MC). In this work, we implemented a PPIAs, starting from ProteinPhosphatase 1 (PP1) expression up to the validation with samples of algal blooms fromArgentina. To do this, we optimized the expression and lyophilization of PP1, and the assayconditions. Also, we included robustness and possible interfering analysis. We evaluated themost widely used cyanobacterial lysis methods and determined that heating for 15 minutes at 95°C is simple and adequate for this assay. Then, we performed MC spikes recoveryassays on water samples from three dams from Argentina, resulting in a recovery rangingfrom 77 to 115%. The limit of detection (LOD) was 0.4 μg/L and the linear range is 0.4 μg/L -5 μg/L. Finally, we evaluated 64 environmental samples where MC was measured by ELISAtest containing from 0 μg/L to 625 μg/L. The PPIA showed excellent correlation (Pearsoncorrelation coefficient = 0.967), no false negative and no false positives above the 1 μg/LWHO guideline (0.11 false positive rate). In conclusion, we optimized and validated a PPIAsto be an effective and accessible alternative to available commercial tests.
Fil: Alba Posse, Jorge Ezequiel. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gonzalez, Carolina. Aguas y Saneamientos Argentinos; Argentina
Fil: Carriquiriborde, Pedro. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigaciones del Medio Ambiente - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones del Medio Ambiente; Argentina
Fil: Nadra, Alejandro Daniel. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gasulla, Javier. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
MICORCYSTIN
CYANOHABS
CYANOTOXINS
PHOSPHATASE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/218157
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/218157
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detectionAlba Posse, Jorge EzequielGonzalez, CarolinaCarriquiriborde, PedroNadra, Alejandro DanielGasulla, JavierMICORCYSTINCYANOHABSCYANOTOXINSPHOSPHATASEhttps://purl.org/becyt/ford/2.8https://purl.org/becyt/ford/2The presence of cyanobacterial toxins in freshwater constitutes an increasing public healthconcern, especially affecting developing countries where the high cost of available methodsmakes monitoring programs difficult. The phosphatase inhibition assay (PPIAs) is a sensitivemethod with low instrument requirements that allows the quantification of the most frequentcyanotoxins, microcystins (MC). In this work, we implemented a PPIAs, starting from ProteinPhosphatase 1 (PP1) expression up to the validation with samples of algal blooms fromArgentina. To do this, we optimized the expression and lyophilization of PP1, and the assayconditions. Also, we included robustness and possible interfering analysis. We evaluated themost widely used cyanobacterial lysis methods and determined that heating for 15 minutes at 95°C is simple and adequate for this assay. Then, we performed MC spikes recoveryassays on water samples from three dams from Argentina, resulting in a recovery rangingfrom 77 to 115%. The limit of detection (LOD) was 0.4 μg/L and the linear range is 0.4 μg/L -5 μg/L. Finally, we evaluated 64 environmental samples where MC was measured by ELISAtest containing from 0 μg/L to 625 μg/L. The PPIA showed excellent correlation (Pearsoncorrelation coefficient = 0.967), no false negative and no false positives above the 1 μg/LWHO guideline (0.11 false positive rate). In conclusion, we optimized and validated a PPIAsto be an effective and accessible alternative to available commercial tests.Fil: Alba Posse, Jorge Ezequiel. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gonzalez, Carolina. Aguas y Saneamientos Argentinos; ArgentinaFil: Carriquiriborde, Pedro. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigaciones del Medio Ambiente - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones del Medio Ambiente; ArgentinaFil: Nadra, Alejandro Daniel. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gasulla, Javier. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaCold Spring Harbor Laboratory Press2022-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/218157Alba Posse, Jorge Ezequiel; Gonzalez, Carolina; Carriquiriborde, Pedro; Nadra, Alejandro Daniel; Gasulla, Javier; Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection; Cold Spring Harbor Laboratory Press; BioRxiv; 2022; 8-2022; 1-392692-8205CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1101/2022.08.16.502937info:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2022.08.16.502937v1info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:43:30Zoai:ri.conicet.gov.ar:11336/218157instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:43:31.126CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection
title Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection
spellingShingle Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection
Alba Posse, Jorge Ezequiel
MICORCYSTIN
CYANOHABS
CYANOTOXINS
PHOSPHATASE
title_short Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection
title_full Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection
title_fullStr Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection
title_full_unstemmed Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection
title_sort Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection
dc.creator.none.fl_str_mv Alba Posse, Jorge Ezequiel
Gonzalez, Carolina
Carriquiriborde, Pedro
Nadra, Alejandro Daniel
Gasulla, Javier
author Alba Posse, Jorge Ezequiel
author_facet Alba Posse, Jorge Ezequiel
Gonzalez, Carolina
Carriquiriborde, Pedro
Nadra, Alejandro Daniel
Gasulla, Javier
author_role author
author2 Gonzalez, Carolina
Carriquiriborde, Pedro
Nadra, Alejandro Daniel
Gasulla, Javier
author2_role author
author
author
author
dc.subject.none.fl_str_mv MICORCYSTIN
CYANOHABS
CYANOTOXINS
PHOSPHATASE
topic MICORCYSTIN
CYANOHABS
CYANOTOXINS
PHOSPHATASE
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.8
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv The presence of cyanobacterial toxins in freshwater constitutes an increasing public healthconcern, especially affecting developing countries where the high cost of available methodsmakes monitoring programs difficult. The phosphatase inhibition assay (PPIAs) is a sensitivemethod with low instrument requirements that allows the quantification of the most frequentcyanotoxins, microcystins (MC). In this work, we implemented a PPIAs, starting from ProteinPhosphatase 1 (PP1) expression up to the validation with samples of algal blooms fromArgentina. To do this, we optimized the expression and lyophilization of PP1, and the assayconditions. Also, we included robustness and possible interfering analysis. We evaluated themost widely used cyanobacterial lysis methods and determined that heating for 15 minutes at 95°C is simple and adequate for this assay. Then, we performed MC spikes recoveryassays on water samples from three dams from Argentina, resulting in a recovery rangingfrom 77 to 115%. The limit of detection (LOD) was 0.4 μg/L and the linear range is 0.4 μg/L -5 μg/L. Finally, we evaluated 64 environmental samples where MC was measured by ELISAtest containing from 0 μg/L to 625 μg/L. The PPIA showed excellent correlation (Pearsoncorrelation coefficient = 0.967), no false negative and no false positives above the 1 μg/LWHO guideline (0.11 false positive rate). In conclusion, we optimized and validated a PPIAsto be an effective and accessible alternative to available commercial tests.
Fil: Alba Posse, Jorge Ezequiel. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gonzalez, Carolina. Aguas y Saneamientos Argentinos; Argentina
Fil: Carriquiriborde, Pedro. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigaciones del Medio Ambiente - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigaciones del Medio Ambiente; Argentina
Fil: Nadra, Alejandro Daniel. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gasulla, Javier. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biociencias, Biotecnología y Biología Traslacional.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description The presence of cyanobacterial toxins in freshwater constitutes an increasing public healthconcern, especially affecting developing countries where the high cost of available methodsmakes monitoring programs difficult. The phosphatase inhibition assay (PPIAs) is a sensitivemethod with low instrument requirements that allows the quantification of the most frequentcyanotoxins, microcystins (MC). In this work, we implemented a PPIAs, starting from ProteinPhosphatase 1 (PP1) expression up to the validation with samples of algal blooms fromArgentina. To do this, we optimized the expression and lyophilization of PP1, and the assayconditions. Also, we included robustness and possible interfering analysis. We evaluated themost widely used cyanobacterial lysis methods and determined that heating for 15 minutes at 95°C is simple and adequate for this assay. Then, we performed MC spikes recoveryassays on water samples from three dams from Argentina, resulting in a recovery rangingfrom 77 to 115%. The limit of detection (LOD) was 0.4 μg/L and the linear range is 0.4 μg/L -5 μg/L. Finally, we evaluated 64 environmental samples where MC was measured by ELISAtest containing from 0 μg/L to 625 μg/L. The PPIA showed excellent correlation (Pearsoncorrelation coefficient = 0.967), no false negative and no false positives above the 1 μg/LWHO guideline (0.11 false positive rate). In conclusion, we optimized and validated a PPIAsto be an effective and accessible alternative to available commercial tests.
publishDate 2022
dc.date.none.fl_str_mv 2022-08
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/218157
Alba Posse, Jorge Ezequiel; Gonzalez, Carolina; Carriquiriborde, Pedro; Nadra, Alejandro Daniel; Gasulla, Javier; Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection; Cold Spring Harbor Laboratory Press; BioRxiv; 2022; 8-2022; 1-39
2692-8205
CONICET Digital
CONICET
url http://hdl.handle.net/11336/218157
identifier_str_mv Alba Posse, Jorge Ezequiel; Gonzalez, Carolina; Carriquiriborde, Pedro; Nadra, Alejandro Daniel; Gasulla, Javier; Optimization and validation of a Protein Phosphatase inhibition assay for accessible microcystin detection; Cold Spring Harbor Laboratory Press; BioRxiv; 2022; 8-2022; 1-39
2692-8205
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1101/2022.08.16.502937
info:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2022.08.16.502937v1
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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