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Specific Six-Transmembrane Epithelial Antigen of the Prostate 1 Capture with Gellan Gum Microspheres

2023-01-18Artigo científico Acesso aberto
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dc.contributor.authorBatista-Silva, João
dc.contributor.authorGomes, Diana
dc.contributor.authorBarroca-Ferreira, Jorge
dc.contributor.authorGallardo, Eugénia
dc.contributor.authorSousa, Ângela
dc.contributor.authorPassarinha, Luís A.
dc.contributor.institutionUCIBIO - Applied Molecular Biosciences Unit
dc.contributor.institutionDQ - Departamento de Química
dc.contributor.pblMDPI - Multidisciplinary Digital Publishing Institute
dc.date.accessioned2023-07-04T22:17:07Z
dc.date.available2023-07-04T22:17:07Z
dc.date.issued2023-01-18
dc.descriptionFunding Information: The authors acknowledge the support from FEDER funds through the POCI-COMPETE 2020–Operational Programme Competitiveness and Internationalisation in Axis I–Strengthening Research, Technological Development and Innovation (Project POCI-01-0145-FEDER-007491).This work was also supported by the Associate Laboratory Institute for Health and Bioeconomy–i4HB (project LA/P/0140/2020) which are financed by National Funds from FCT/MCTES. Publisher Copyright: © 2023 by the authors.
dc.description.abstractThis work demonstrates the potential of calcium- and nickel-crosslinked Gellan Gum (GG) microspheres to capture the Six-Transmembrane Epithelial Antigen of the Prostate 1 (STEAP1) directly from complex Komagataella pastoris mini-bioreactor lysates in a batch method. Calcium-crosslinked microspheres were applied in an ionic exchange strategy, by manipulation of pH and ionic strength, whereas nickel-crosslinked microspheres were applied in an affinity strategy, mirroring a standard immobilized metal affinity chromatography. Both formulations presented small diameters, with appreciable crosslinker content, but calcium-crosslinked microspheres were far smoother. The most promising results were obtained for the ionic strategy, wherein calcium-crosslinked GG microspheres were able to completely bind 0.1% (v/v) DM solubilized STEAP1 in lysate samples (~7 mg/mL). The target protein was eluted in a complexed state at pH 11 with 500 mM NaCl in 10 mM Tris buffer, in a single step with minimal losses. Coupling the batch clarified sample with a co-immunoprecipitation polishing step yields a sample of monomeric STEAP1 with a high degree of purity. For the first time, we demonstrate the potential of a gellan batch method to function as a clarification and primary capture method towards STEAP1, a membrane protein, simplifying and reducing the costs of standard purification workflows.en
dc.description.versionpublishersversion
dc.description.versionpublished
dc.format.extent19
dc.format.extent3102676
dc.identifier.doi10.3390/ijms24031949
dc.identifier.issn1661-6596
dc.identifier.otherPURE: 65198713
dc.identifier.otherPURE UUID: 590a37f1-7cc6-460d-a531-7d93bc57e814
dc.identifier.otherScopus: 85147895711
dc.identifier.otherWOS: 000929578200001
dc.identifier.otherPubMed: 36768273
dc.identifier.otherPubMedCentral: PMC9916199
dc.identifier.urihttp://hdl.handle.net/10362/154840
dc.identifier.urlhttps://www.scopus.com/pages/publications/85147895711
dc.language.isoeng
dc.peerreviewedyes
dc.relationinfo:eu-repo/grantAgreement/FCT/OE/SFRH%2FBD%2F130068%2F2017/PT
dc.relationStructural and Functional Characterization of the Cancer-associated Epithelial Antigen STEAP1
dc.relationBiointeration studies of recombinant HPV-16 oncoproteins to discover therapeutic inhibitors
dc.relationHealth Sciences Research Centre
dc.relationHealth Sciences Research Centre
dc.relationApplied Molecular Biosciences Unit
dc.relationApplied Molecular Biosciences Unit
dc.relationinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT
dc.relationinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT
dc.subjectbatch method
dc.subjectcapture
dc.subjectco-immunoprecipitation
dc.subjectGellan gum microspheres
dc.subjectSTEAP1
dc.subjectCatalysis
dc.subjectMolecular Biology
dc.subjectSpectroscopy
dc.subjectComputer Science Applications
dc.subjectPhysical and Theoretical Chemistry
dc.subjectOrganic Chemistry
dc.subjectInorganic Chemistry
dc.titleSpecific Six-Transmembrane Epithelial Antigen of the Prostate 1 Capture with Gellan Gum Microspheresen
dc.title.subtitleDesign, Optimization and Integrationen
dc.typejournal article
degois.publication.issue3
degois.publication.titleInternational Journal of Molecular Sciences
degois.publication.volume24
dspace.entity.typePublication
oaire.awardNumberSFRH/BD/130068/2017
oaire.awardNumber2020.06792.BD
oaire.awardNumberUIDB/00709/2020
oaire.awardNumberUIDP/00709/2020
oaire.awardNumberUIDB/04378/2020
oaire.awardNumberUIDP/04378/2020
oaire.awardTitleStructural and Functional Characterization of the Cancer-associated Epithelial Antigen STEAP1
oaire.awardTitleBiointeration studies of recombinant HPV-16 oncoproteins to discover therapeutic inhibitors
oaire.awardTitleHealth Sciences Research Centre
oaire.awardTitleHealth Sciences Research Centre
oaire.awardTitleApplied Molecular Biosciences Unit
oaire.awardTitleApplied Molecular Biosciences Unit
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/OE/SFRH%2FBD%2F130068%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//2020.06792.BD/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F00709%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F00709%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT
oaire.fundingStreamOE
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
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