Repository logo
 
Publication

Assessment of immunoglobulin capture in immobilized protein A through automatic bead injection

2019Journal article Restricted access
Simple item page
dc.contributor.authorRamos, Inês I.
dc.contributor.authorMarques, Sara S.
dc.contributor.authorMagalhães, Luís M.
dc.contributor.authorBarreiros, Luisa
dc.contributor.authorReis, Salette
dc.contributor.authorLima, José L.F. C.
dc.contributor.authorSegundo, Marcela A.
dc.date.accessioned2020-09-14T13:07:59Z
dc.date.available2020-09-14T13:07:59Z
dc.date.issued2019
dc.description.abstractThe repeatable immobilization of molecular recognition elements onto particle surfaces has a strong impact on the outcomes of affinity-based assays. In this work, an automatic method for the immobilization of immunoglobulin G (IgG) onto protein A-Sepharose microbeads was established through the flow programming features of the portable lab-on-valve platform using micro-bead injection spectroscopy. The reproducible packing of protein A-microbeads between two optic fibers was feasible, allowing on-column probing of IgG retention. The automation of solutions handling and the precise control of time of IgG interaction with the beads rendered repeatable immobilization cycles, within a short timeframe (<2 min). The proposed method featured the preparation of disposable immunosorbents for downstream analytical applications, such as immunosensing or microenrichment of target analytes. In-situ quantification of IgG@protein A-microbeads was carried out using a horseradish peroxidase-labeled detection IgG. The colorimetric oxidation of 3,3',5,5'-tetramethylbenzidine was monitored on-column. Quantitation of mouse and human IgG immobilized@protein A-microbeads was achieved for loading masses between 0.1 and 0.4 μg per ca. 5.5 mg of sorbent. The implemented detection strategy allowed the quantification of human IgG in certified human serum (ERM®- DA470k/IFCC) and spiked saliva, yielding recoveries of 102-108% and requiring minimal volume (1-15 μL) from serum and saliva.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.doi10.1016/j.talanta.2019.06.023pt_PT
dc.identifier.urihttp://hdl.handle.net/10400.22/16263
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.relationPOCI/01/0145/FEDER/007265pt_PT
dc.relationNanodiagnostics and nanotherapeutics
dc.relationAssociated Laboratory for Green Chemistry - Clean Technologies and Processes
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S0039914019306484pt_PT
dc.subjectArmoraciapt_PT
dc.subjectBenzidinespt_PT
dc.subjectChromatography, Affinitypt_PT
dc.subjectChromogenic Compoundspt_PT
dc.subjectColorimetrypt_PT
dc.subjectHorseradish Peroxidasept_PT
dc.subjectImmobilized Proteinspt_PT
dc.subjectImmunoglobulin Gpt_PT
dc.subjectMicrospherespt_PT
dc.subjectOxidation-Reductionpt_PT
dc.subjectSalivapt_PT
dc.subjectSepharosept_PT
dc.subjectStaphylococcal Protein Apt_PT
dc.titleAssessment of immunoglobulin capture in immobilized protein A through automatic bead injectionpt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleNanodiagnostics and nanotherapeutics
oaire.awardTitleAssociated Laboratory for Green Chemistry - Clean Technologies and Processes
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBD%2F97540%2F2013/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//PD%2FBI%2F113970%2F2015/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UID%2FQUI%2F50006%2F2019/PT
oaire.citation.endPage547pt_PT
oaire.citation.startPage542pt_PT
oaire.citation.titleTalantapt_PT
oaire.citation.volume204pt_PT
oaire.fundingStreamSFRH
oaire.fundingStream6817 - DCRRNI ID
person.familyNameBarreiros
person.givenNameLuisa
person.identifier.ciencia-id611F-E0C5-0230
person.identifier.orcid0000-0003-3481-5809
person.identifier.ridD-7950-2013
person.identifier.scopus-author-id6508205485
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
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication1e66bacc-64de-4ecb-96b7-4c0e366cba57
relation.isAuthorOfPublication.latestForDiscovery1e66bacc-64de-4ecb-96b7-4c0e366cba57
relation.isProjectOfPublication03b667da-b739-46fb-986e-5c919289a656
relation.isProjectOfPublicationc364a358-efe4-4386-944b-87cd3e4643f8
relation.isProjectOfPublicationf943d798-b7fe-4c3c-87a7-46b3e5ac86be
relation.isProjectOfPublication.latestForDiscoveryf943d798-b7fe-4c3c-87a7-46b3e5ac86be

Files

Original bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
ART_LuísaBarreiros_2019.pdf
Size:
776.59 KB
Format:
Adobe Portable Document Format
Download

Collections

ESS - FAR - Artigos