Repository logo
 
Publication

Refinement of two-dimensional electrophoresis for vitreous proteome profiling using an artificial neural network

2019Journal article Open access
Simple item page
dc.contributor.authorSantos, Fátima Raquel Milhano dos
dc.contributor.authorAlbuquerque, Tânia Gonçalves
dc.contributor.authorGaspar, Leonor Isabel Mesquita
dc.contributor.authorDias, Joao ML
dc.contributor.authorSousa, João Paulo Castro De
dc.contributor.authorParadela, Alberto
dc.contributor.authorTomaz, C. T.
dc.contributor.authorPassarinha, LA
dc.date.accessioned2020-02-11T16:27:34Z
dc.date.available2022-01-01T01:30:10Z
dc.date.issued2019
dc.description.abstractDespite technological advances, two-dimensional electrophoresis (2DE) of biological fluids, such as vitreous, remains a major challenge. In this study, artificial neural network was applied to optimize the recovery of vitreous proteins and its detection by 2DE analysis through the combination of several solubilizing agents (CHAPS, Genapol, DTT, IPG buffer), temperature, and total voltage. The highest protein recovery (94.9% ± 4.5) was achieved using 4% (w/v) CHAPS, 0.1% (v/v) Genapol, 20 mM DTT, and 2% (v/v) IPG buffer. Two iterations were required to achieve an optimized response (580 spots) using 4% (w/v) CHAPS, 0.2% (v/v) Genapol, 60 mM DTT, and 0.5% (v/v) IPG buffer at 35 kVh and 25 °C, representing a 2.4-fold improvement over the standard initial conditions of the experimental design. The analysis of depleted vitreous using the optimized protocol resulted in an additional 1.3-fold increment in protein detection over the optimal output, with an average of 761 spots detected in vitreous from different vitreoretinopathies. Our results clearly indicate the importance of combining the appropriate amount of solubilizing agents with a suitable control of the temperature and voltage to obtain high-quality gels. The high-throughput of this model provides an effective starting point for the optimization of 2DE protocols. This experimental design can be adapted to other types of matrices. Graphical abstract.pt_PT
dc.description.sponsorshipCENTRO-07-ST24-FEDER-002014; CNB-CSIC is supported by grant PT13/0001, of the PE I +D+i 2013–2016, funded by ISCIII and FEDER.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1007/s00216-019-01887-ypt_PT
dc.identifier.urihttp://hdl.handle.net/10400.6/9211
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherSpringerpt_PT
dc.relationVitreous humor biomarkers in Age-related Macular Degeneration
dc.relationApplied Molecular Biosciences Unit
dc.relationHealth Sciences Research Centre
dc.subjectTwo-Dimensional gel electrophoresispt_PT
dc.subjectGel-based proteomicspt_PT
dc.subjectArtificial Neural Networkspt_PT
dc.subjectOcular pathologiespt_PT
dc.subjectVitreouspt_PT
dc.titleRefinement of two-dimensional electrophoresis for vitreous proteome profiling using an artificial neural networkpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleVitreous humor biomarkers in Age-related Macular Degeneration
oaire.awardTitleApplied Molecular Biosciences Unit
oaire.awardTitleHealth Sciences Research Centre
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/POR_CENTRO/SFRH%2FBD%2F112526%2F2015/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/5876/UID%2FMulti%2F00709%2F2013/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UID%2FMulti%2F04378%2F2019/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F00709%2F2020/PT
oaire.citation.endPage5126pt_PT
oaire.citation.issue20pt_PT
oaire.citation.startPage5115pt_PT
oaire.citation.titleAnalytical and Bioanalytical Chemistrypt_PT
oaire.citation.volume411pt_PT
oaire.fundingStreamPOR_CENTRO
oaire.fundingStream5876
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
person.familyNameMilhano dos Santos
person.familyNameGaspar
person.familyNameDias
person.familyNameCastro de Sousa
person.familyNameParadela
person.familyNameTomaz
person.familyNamePaulino Passarinha
person.givenNameFátima Raquel
person.givenNameLeonor Isabel Mesquita
person.givenNameJoao
person.givenNameJoão Paulo
person.givenNameAlberto
person.givenNameCândida
person.givenNameLuís António
person.identifier2198714
person.identifierhttps://www.researchgate.net/profile/Luis_Passarinha
person.identifier.ciencia-idD01A-2666-0E20
person.identifier.ciencia-id3216-14E9-65E0
person.identifier.ciencia-id191F-1685-A755
person.identifier.ciencia-idD81B-D3DE-23D2
person.identifier.ciencia-idFE13-0C44-13D1
person.identifier.ciencia-id621B-74BF-F9A7
person.identifier.orcid0000-0002-4041-1504
person.identifier.orcid0000-0002-5106-1888
person.identifier.orcid0000-0002-8451-3537
person.identifier.orcid0000-0002-0025-1841
person.identifier.orcid0000-0001-6837-7056
person.identifier.orcid0000-0002-0927-2331
person.identifier.orcid0000-0001-6910-7576
person.identifier.ridD-2011-2013
person.identifier.ridH-2113-2015
person.identifier.ridF-9367-2014
person.identifier.scopus-author-id56262249200
person.identifier.scopus-author-id6602585990
person.identifier.scopus-author-id7004546303
person.identifier.scopus-author-id12445439600
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.embargofctCopyright cedido à editora no momento da publicaçãopt_PT
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication02084069-54f0-41a2-80a1-9edd01bb5895
relation.isAuthorOfPublication00904288-9953-4e68-ad3e-6fad566f2845
relation.isAuthorOfPublication8a4bd905-38bd-4d4f-88d3-d542ad3e365d
relation.isAuthorOfPublicationb53f642a-416d-4873-a962-3c1b34306e6f
relation.isAuthorOfPublication465880c9-b09c-436f-a69d-9f178b2d688d
relation.isAuthorOfPublicationa8b816a2-a48c-43b1-a064-2ae8aa4b9bf8
relation.isAuthorOfPublicationa431a31c-1ae0-43e9-8d84-4e9caca2ad92
relation.isAuthorOfPublication.latestForDiscoveryb53f642a-416d-4873-a962-3c1b34306e6f
relation.isProjectOfPublicationb843a31e-e5b1-4b4d-8944-a74f8a82f84c
relation.isProjectOfPublicationcb4688cf-40da-4197-9eb0-c55bad699ad3
relation.isProjectOfPublicationdcece67e-f19f-4f30-b8b3-cfa5605a4025
relation.isProjectOfPublicationc7cee677-067c-47b2-ab19-b2dd5b0fda9d
relation.isProjectOfPublication.latestForDiscoverycb4688cf-40da-4197-9eb0-c55bad699ad3

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
44Santos2019_Article_Refinement2D_AnalBioChem.pdf
Size:
4.63 MB
Format:
Adobe Portable Document Format
Description:
Article_Refinement2D_AnalBioChem
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.71 KB
Format:
Item-specific license agreed upon to submission
Description:
Download

Collections

ICI - CICS | Documentos por Auto-Depósito
Centros - UBIMEDICAL | Documentos por Auto-Depósito
FC - DQ | Documentos por Auto-Depósito
FCS - DCM | Documentos por Auto-Depósito