Modeling and simulation of anion exchange chromatography for purification of proteins in complex mixtures

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dc.contributorLEDV - Laboratório de Desenvolvimento de Vacinaspt_BR
dc.contributor.authorBenedini, Leandro J.pt_BR
dc.contributor.authorFigueiredo, Douglas Borgespt_BR
dc.contributor.authorCabrera-Crespo, Joaquinpt_BR
dc.contributor.authorGonçalves, Viviane Maimonipt_BR
dc.contributor.authorSilva, Gabriel G.pt_BR
dc.contributor.authorCampani, Gilsonpt_BR
dc.contributor.authorZangirolami, Teresa C.pt_BR
dc.contributor.authorFurlan, Felipe F.pt_BR
dc.date.accessioned2020-07-09T21:26:59Z-
dc.date.available2020-07-09T21:26:59Z-
dc.date.issued2020pt_BR
dc.identifier.citationBenedini LJ., Figueiredo DB, Cabrera-Crespo J, Gonçalves VM, Silva GG., Campani G, et al. Modeling and simulation of anion exchange chromatography for purification of proteins in complex mixtures. J. Chromatogr. A. 2020 Feb;1613:460685. doi:10.1016/j.chroma.2019.460685.pt_BR
dc.identifier.urihttps://repositorio.butantan.gov.br/handle/butantan/2980-
dc.description.abstractIon exchange chromatography is extensively used in the purification of biological compounds. Reliable mathematical models describing this chromatographic technique are available and can be used to improve the performance of this separation step. However, the use of synthetic mixtures for model development hampers the application of this approach with real cell extracts processed in downstream operations. This work presents an original approach for handling non-synthetic genuine mixtures of proteins, which was applied in the purification of an untagged recombinant pneumococcal surface protein A (PspA4Pro). First, evaluation was made of the efficiency of steric mass action (SMA) and modified Langmuir isotherms, which were separately used together with the equilibrium dispersive model (EDM). The data used for parameter estimation and model validation were obtained from anion exchange chromatography runs (employing Q-Sepharose FF), applied to real cell extracts produced by different cultivation strategies. Simulations showed that the models were able to describe the complex mixtures of unknown proteins. Next, the EDM and SMA approaches were used to separately describe the profile of PspA4Pro and the pool of protein impurities eluted together. The simulations showed that PspA4Pro tended to elute at the beginning of the peak, enabling the establishment of an alternative elution schedule that provided a 34% increase in the purity achieved using the anion exchange chromatography.pt_BR
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)pt_BR
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)pt_BR
dc.format.extent460685pt_BR
dc.language.isoEnglishpt_BR
dc.relation.ispartofJournal of Chromatography Apt_BR
dc.titleModeling and simulation of anion exchange chromatography for purification of proteins in complex mixturespt_BR
dc.typeArticlept_BR
dc.identifier.doi10.1016/j.chroma.2019.460685pt_BR
dc.identifier.urlhttps://doi.org/10.1016/j.chroma.2019.460685pt_BR
dc.contributor.externalUniversidade Federal de São Carlos (UFSCar)¦¦Brasilpt_BR
dc.contributor.externalInstituto Federal de São Paulo (IFSP)¦¦Brasilpt_BR
dc.contributor.externalUniversidade Federal de Lavras (UFLA)¦¦Brasilpt_BR
dc.identifier.citationvolume1613pt_BR
dc.subject.keywordProtein purificationpt_BR
dc.subject.keywordAnion exchange chromatographypt_BR
dc.subject.keywordMathematical modelingpt_BR
dc.subject.keywordComplex protein mixturept_BR
dc.subject.keywordSteric mass action isothermpt_BR
dc.subject.keywordEquilibrium dispersive modelpt_BR
dc.relation.ispartofabbreviatedJ Chromatogr Apt_BR
dc.identifier.citationabntv. 1613, 460685, fev. 2020pt_BR
dc.identifier.citationvancouver2020 Feb;1613:460685pt_BR
dc.contributor.butantanFigueiredo, Douglas Borges|:Aluno|:LEDV - Laboratório de Desenvolvimento de Vacinas|:pt_BR
dc.contributor.butantanCabrera-Crespo, Joaquin|:Pesquisador|:LEDV - Laboratório de Desenvolvimento de Vacinas|:pt_BR
dc.contributor.butantanGonçalves, Viviane Maimoni|:Pesquisador|:LEDV - Laboratório de Desenvolvimento de Vacinas|:pt_BR
dc.sponsorship.butantanCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)¦¦001pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2015/10291pt_BR
dc.identifier.bvsccBR78.1pt_BR
dc.identifier.bvsdbIBProdpt_BR
dc.description.dbindexedYespt_BR
item.grantfulltextembargo_29990101-
item.languageiso639-1English-
item.openairetypeArticle-
item.fulltextCom Texto completo-
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