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In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy

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Resumo:BACKGROUNDWhile the pharmaceutical industry keeps an eye on plasmid DNA production for new generation gene therapies, real-time monitoring techniques for plasmid bioproduction are as yet unavailable. This work shows the possibility of in situ monitoring of plasmid production in Escherichia coli cultures using a near infrared (NIR) fiber optic probe. RESULTSPartial least squares (PLS) regression models based on the NIR spectra were developed for predicting bioprocess critical variables such as the concentrations of biomass, plasmid, carbon sources (glucose and glycerol) and acetate. In order to achieve robust models able to predict the performance of plasmid production processes, independently of the composition of the cultivation medium, cultivation strategy (batch versus fed-batch) and E. coli strain used, three strategies were adopted, using: (i) E. coliDH5 cultures conducted under different media compositions and culture strategies (batch and fed-batch); (ii) engineered E. coli strains, MG1655endArecApgi and MG1655endArecA, grown on the same medium and culture strategy; (iii) diverse E. coli strains, over batch and fed-batch cultivations and using different media compositions. PLS models showed high accuracy for predicting all variables in the three groups of cultures. CONCLUSIONNIR spectroscopy combined with PLS modeling provides a fast, inexpensive and contamination-free technique to accurately monitoring plasmid bioprocesses in real time, independently of the medium composition, cultivation strategy and the E. coli strain used.
Autores principais:Lopes, Marta B.
Outros Autores:Gonçalves, Geisa A. L.; Felício-Silva, Daniel; Prather, Kristala L. J.; Monteiro, Gabriel; Prazeres, Duarte M. F.; Calado, Cecília
Assunto:Plasmid bioprocess Escherichia coli Near infrared spectroscopy In situ monitoring
Ano:2015
País:Portugal
Tipo de documento:artigo
Tipo de acesso:acesso restrito
Instituição associada:Instituto Politécnico de Lisboa
Idioma:inglês
Origem:Repositório Científico do Instituto Politécnico de Lisboa
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author Lopes, Marta B.
author2 Gonçalves, Geisa A. L.
Felício-Silva, Daniel
Prather, Kristala L. J.
Monteiro, Gabriel
Prazeres, Duarte M. F.
Calado, Cecília
author2_role author
author
author
author
author
author
author_facet Lopes, Marta B.
Gonçalves, Geisa A. L.
Felício-Silva, Daniel
Prather, Kristala L. J.
Monteiro, Gabriel
Prazeres, Duarte M. F.
Calado, Cecília
author_role author
contributor_name_str_mv RCIPL
country_str PT
creators_json_txt [{\"Person.name\":\"Lopes, Marta B.\"},{\"Person.name\":\"Gonçalves, Geisa A. L.\"},{\"Person.name\":\"Felício-Silva, Daniel\"},{\"Person.name\":\"Prather, Kristala L. J.\"},{\"Person.name\":\"Monteiro, Gabriel\"},{\"Person.name\":\"Prazeres, Duarte M. F.\"},{\"Person.name\":\"Calado, Cecília\",\"Person.identifier.orcid\":\"0000-0002-5264-9755\"}]
datacite.contributors.contributor.contributorName.fl_str_mv RCIPL
datacite.creators.creator.creatorName.fl_str_mv Lopes, Marta B.
Gonçalves, Geisa A. L.
Felício-Silva, Daniel
Prather, Kristala L. J.
Monteiro, Gabriel
Prazeres, Duarte M. F.
Calado, Cecília
datacite.date.Accepted.fl_str_mv 2015-02-01T00:00:00Z
datacite.date.available.fl_str_mv 2016-04-20T14:20:47Z
datacite.date.embargoed.fl_str_mv 2016-04-20T14:20:47Z
datacite.rights.fl_str_mv http://purl.org/coar/access_right/c_16ec
datacite.subjects.subject.fl_str_mv Plasmid bioprocess
Escherichia coli
Near infrared spectroscopy
In situ monitoring
datacite.titles.title.fl_str_mv In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy
dc.contributor.none.fl_str_mv RCIPL
dc.creator.none.fl_str_mv Lopes, Marta B.
Gonçalves, Geisa A. L.
Felício-Silva, Daniel
Prather, Kristala L. J.
Monteiro, Gabriel
Prazeres, Duarte M. F.
Calado, Cecília
dc.date.Accepted.fl_str_mv 2015-02-01T00:00:00Z
dc.date.available.fl_str_mv 2016-04-20T14:20:47Z
dc.date.embargoed.fl_str_mv 2016-04-20T14:20:47Z
dc.format.none.fl_str_mv application/pdf
dc.identifier.none.fl_str_mv http://hdl.handle.net/10400.21/6058
dc.language.none.fl_str_mv eng
dc.publisher.none.fl_str_mv Wiley-Blackwell
dc.rights.none.fl_str_mv http://purl.org/coar/access_right/c_16ec
dc.subject.none.fl_str_mv Plasmid bioprocess
Escherichia coli
Near infrared spectroscopy
In situ monitoring
dc.title.fl_str_mv In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy
dc.type.none.fl_str_mv http://purl.org/coar/resource_type/c_6501
description BACKGROUNDWhile the pharmaceutical industry keeps an eye on plasmid DNA production for new generation gene therapies, real-time monitoring techniques for plasmid bioproduction are as yet unavailable. This work shows the possibility of in situ monitoring of plasmid production in Escherichia coli cultures using a near infrared (NIR) fiber optic probe. RESULTSPartial least squares (PLS) regression models based on the NIR spectra were developed for predicting bioprocess critical variables such as the concentrations of biomass, plasmid, carbon sources (glucose and glycerol) and acetate. In order to achieve robust models able to predict the performance of plasmid production processes, independently of the composition of the cultivation medium, cultivation strategy (batch versus fed-batch) and E. coli strain used, three strategies were adopted, using: (i) E. coliDH5 cultures conducted under different media compositions and culture strategies (batch and fed-batch); (ii) engineered E. coli strains, MG1655endArecApgi and MG1655endArecA, grown on the same medium and culture strategy; (iii) diverse E. coli strains, over batch and fed-batch cultivations and using different media compositions. PLS models showed high accuracy for predicting all variables in the three groups of cultures. CONCLUSIONNIR spectroscopy combined with PLS modeling provides a fast, inexpensive and contamination-free technique to accurately monitoring plasmid bioprocesses in real time, independently of the medium composition, cultivation strategy and the E. coli strain used.
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fulltext.url.fl_str_mv https://repositorio.ipl.pt/bitstreams/553f4498-f213-4444-9cc4-bb3df9a67815/download
funding.funder.alternateName_str_mv FCT
funding.funder.identifier_str_mv http://doi.org/10.13039/501100001871
funding.funder.name_str_mv Fundação para a Ciência e a Tecnologia
id ripl_c135d7ea7f9cdc8a528994ded96843fd
identifier.url.fl_str_mv http://hdl.handle.net/10400.21/6058
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instname_str Instituto Politécnico de Lisboa
language eng
network_acronym_str ripl
network_name_str Repositório Científico do Instituto Politécnico de Lisboa
oai_identifier_str oai:repositorio.ipl.pt:10400.21/6058
organization_str_mv urn:organizationAcronym:ipl
person_str_mv Lopes, Marta B.
Gonçalves, Geisa A. L.
Felício-Silva, Daniel
Prather, Kristala L. J.
Monteiro, Gabriel
Prazeres, Duarte M. F.
Calado, Cecília
Calado, Cecília
https://www.ciencia-id.pt/9418-E320-3177
9418-E320-3177
http://orcid.org/0000-0002-5264-9755
0000-0002-5264-9755
publishDate 2015
publisher.none.fl_str_mv Wiley-Blackwell
reponame_str Repositório Científico do Instituto Politécnico de Lisboa
repository_id_str urn:repositoryAcronym:ripl
service_str_mv urn:repositoryAcronym:ripl
spelling engWiley-Blackwellpt_PTBACKGROUNDWhile the pharmaceutical industry keeps an eye on plasmid DNA production for new generation gene therapies, real-time monitoring techniques for plasmid bioproduction are as yet unavailable. This work shows the possibility of in situ monitoring of plasmid production in Escherichia coli cultures using a near infrared (NIR) fiber optic probe. RESULTSPartial least squares (PLS) regression models based on the NIR spectra were developed for predicting bioprocess critical variables such as the concentrations of biomass, plasmid, carbon sources (glucose and glycerol) and acetate. In order to achieve robust models able to predict the performance of plasmid production processes, independently of the composition of the cultivation medium, cultivation strategy (batch versus fed-batch) and E. coli strain used, three strategies were adopted, using: (i) E. coliDH5 cultures conducted under different media compositions and culture strategies (batch and fed-batch); (ii) engineered E. coli strains, MG1655endArecApgi and MG1655endArecA, grown on the same medium and culture strategy; (iii) diverse E. coli strains, over batch and fed-batch cultivations and using different media compositions. PLS models showed high accuracy for predicting all variables in the three groups of cultures. CONCLUSIONNIR spectroscopy combined with PLS modeling provides a fast, inexpensive and contamination-free technique to accurately monitoring plasmid bioprocesses in real time, independently of the medium composition, cultivation strategy and the E. coli strain used.application/pdfpt_PTIn situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategyLopes, Marta B.Gonçalves, Geisa A. L.Felício-Silva, DanielPrather, Kristala L. J.Monteiro, GabrielPrazeres, Duarte M. F.PersonalCalado, CecíliaDSpacehttp://dspace.org/items/e8577257-c64c-4481-9b2b-940fedb360ccDSpacehttp://dspace.org/items/e8577257-c64c-4481-9b2b-940fedb360ccCaladoCecíliaCiência IDhttps://www.ciencia-id.pt9418-E320-3177ORCIDhttp://orcid.org0000-0002-5264-9755Researcher IDhttps://www.researcherid.comE-2102-2014Researcher IDhttps://www.researcherid.comE-2102-2014Scopus Author IDhttps://www.scopus.com6603163260HostingInstitutionOrganizationalRCIPLe-mailmailto:rcaap@sp.ipl.ptrcaap@sp.ipl.ptISSNIsPartOf0268-2575ISSNIsPartOf1097-4660DOIIsPartOf10.1002/jctb.44312016-04-20T14:20:47Z2015-022015-02-01T00:00:00ZHandlehttp://hdl.handle.net/10400.21/6058http://purl.org/coar/access_right/c_16ecrestricted accessPlasmid bioprocessEscherichia coliNear infrared spectroscopyIn situ monitoring822701 bytesFundação para a Ciência e a TecnologiaRATIONAL ENGINEERING OF E. COLI STRAINS AND VECTORS FOR IMPROVED MANUFACTURING OF PLASMID BIOPHARMACEUTICALSCrossref Funder IDhttp://doi.org/10.13039/501100001871literaturehttp://purl.org/coar/resource_type/c_6501journal articlehttp://purl.org/coar/access_right/c_16ecapplication/pdffulltexthttps://repositorio.ipl.pt/bitstreams/553f4498-f213-4444-9cc4-bb3df9a67815/download902255261
spellingShingle In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy
Lopes, Marta B.
Plasmid bioprocess
Escherichia coli
Near infrared spectroscopy
In situ monitoring
status SINGLETON
subject.fl_str_mv Plasmid bioprocess
Escherichia coli
Near infrared spectroscopy
In situ monitoring
title In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy
title_full In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy
title_fullStr In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy
title_full_unstemmed In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy
title_short In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy
title_sort In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy
topic Plasmid bioprocess
Escherichia coli
Near infrared spectroscopy
In situ monitoring
topic_facet Plasmid bioprocess
Escherichia coli
Near infrared spectroscopy
In situ monitoring
url http://hdl.handle.net/10400.21/6058
visible 1