Pesquisa de métodos cromatográficos para quantificação e caracterização do certolizumabe pegol
Autor(a) principal: | |
---|---|
Data de Publicação: | 2022 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Manancial - Repositório Digital da UFSM |
Texto Completo: | http://repositorio.ufsm.br/handle/1/26640 |
Resumo: | Certolizumab pegol (CZP) is a fragment of a recombinant humanized monoclonal antibody (mAb) produced in E. coli, conjugated with polyethylene glycol (PEG) that acts by inhibiting the activity of Tumor Necrosis Factor alpha (TNF-α). CZP has a therapeutic indication for the treatment of chronic inflammatory diseases such as rheumatoid arthritis and Crohn's disease. Size exclusion chromatography (SEC) and Reversed phase liquid chromatography (RP-LC) methods were developed and validated for quantification of CZP in biotechnological products. The CL-EM method was developed on a BioSep-SEC-S3000 chromatographic column (300 x 4.6 mm, 5μm, 290 Å) maintained at 35°C. Mobile phase A consisted of 100mM monobasic sodium phosphate and 200mM sodium chloride pH 7.0 and mobile phase B consisted of Ethanol (95:5, v/v), with a flow of 0.5mL/min and detection by DAD at 214 nm. In the CL-FR method, the analytical condition was determined with a Zorbax 300SB C18 column (4.6 x 150 mm, 3.5 µm), maintained at 80°C, with mobile phase A consisting of 0.1% (v/v) of trifluoroacetic acid (TFA) in water and mobile phase B prepared by propanol:acetonitrile:water:TFA (70:20:9,9:0.1, v/v). Elution took place in a mobile phase concentration gradient at a flow of 1 ml/min and detection by a diode array detector (DAD) at 214nm. CZP was eluted at retention times of 5.6 and 9.0 min, being linear in the concentration range of 1 - 40 mg / mL (r² = 0.9993) and 1 - 40 mg / mL (r² = 0.9997), respectively, for SEC and for RP-LC. The specificity of the methods was investigated by forced degradation studies, interference of formulation excipients and analysis of peak purity. The limits of detection and quantification were 0.14 and 0.41 mg / mL for the SEC method and 0.06 and 0.17 mg / mL for RP-LC. The accuracy means were 100.50 and 99.80 with a bias of 0.85 and 0.82%, respectively, for the SEC and RP-LC methods. The validation of the methods followed the guidelines established in the official guides of the National Health Surveillance Agency (ANVISA) and the International Conference on Harmonization (ICH). In addition, a cell culture bioassay using the MONO-MAC-6 stimulated by lipopolysaccharide (LPS) was studied to evaluate the biological activity of CZP and inhibition of TNF-α release, which proved to be significant. Correlation studies were carried out between the validated methods, aiming to contribute to improve the quality control of the biotechnological product, in order to ensure safety and therapeutic efficacy. |
id |
UFSM-20_84af1b00651eef2420341e63f61cd055 |
---|---|
oai_identifier_str |
oai:repositorio.ufsm.br:1/26640 |
network_acronym_str |
UFSM-20 |
network_name_str |
Manancial - Repositório Digital da UFSM |
repository_id_str |
3913 |
spelling |
2022-10-24T14:19:08Z2022-10-24T14:19:08Z2022-05-24http://repositorio.ufsm.br/handle/1/26640Certolizumab pegol (CZP) is a fragment of a recombinant humanized monoclonal antibody (mAb) produced in E. coli, conjugated with polyethylene glycol (PEG) that acts by inhibiting the activity of Tumor Necrosis Factor alpha (TNF-α). CZP has a therapeutic indication for the treatment of chronic inflammatory diseases such as rheumatoid arthritis and Crohn's disease. Size exclusion chromatography (SEC) and Reversed phase liquid chromatography (RP-LC) methods were developed and validated for quantification of CZP in biotechnological products. The CL-EM method was developed on a BioSep-SEC-S3000 chromatographic column (300 x 4.6 mm, 5μm, 290 Å) maintained at 35°C. Mobile phase A consisted of 100mM monobasic sodium phosphate and 200mM sodium chloride pH 7.0 and mobile phase B consisted of Ethanol (95:5, v/v), with a flow of 0.5mL/min and detection by DAD at 214 nm. In the CL-FR method, the analytical condition was determined with a Zorbax 300SB C18 column (4.6 x 150 mm, 3.5 µm), maintained at 80°C, with mobile phase A consisting of 0.1% (v/v) of trifluoroacetic acid (TFA) in water and mobile phase B prepared by propanol:acetonitrile:water:TFA (70:20:9,9:0.1, v/v). Elution took place in a mobile phase concentration gradient at a flow of 1 ml/min and detection by a diode array detector (DAD) at 214nm. CZP was eluted at retention times of 5.6 and 9.0 min, being linear in the concentration range of 1 - 40 mg / mL (r² = 0.9993) and 1 - 40 mg / mL (r² = 0.9997), respectively, for SEC and for RP-LC. The specificity of the methods was investigated by forced degradation studies, interference of formulation excipients and analysis of peak purity. The limits of detection and quantification were 0.14 and 0.41 mg / mL for the SEC method and 0.06 and 0.17 mg / mL for RP-LC. The accuracy means were 100.50 and 99.80 with a bias of 0.85 and 0.82%, respectively, for the SEC and RP-LC methods. The validation of the methods followed the guidelines established in the official guides of the National Health Surveillance Agency (ANVISA) and the International Conference on Harmonization (ICH). In addition, a cell culture bioassay using the MONO-MAC-6 stimulated by lipopolysaccharide (LPS) was studied to evaluate the biological activity of CZP and inhibition of TNF-α release, which proved to be significant. Correlation studies were carried out between the validated methods, aiming to contribute to improve the quality control of the biotechnological product, in order to ensure safety and therapeutic efficacy.O certolizumabe pegol (CZP) é um fragmento de um anticorpo monoclonal (mAb) humanizado recombinante produzido em E. coli, conjugado com polietilenoglicol (PEG) que atua inibindo a atividade do Fator de Necrose Tumoral alfa (TNF-α). O CZP apresenta indicação terapêutica no tratamento de enfermidades inflamatórias crônicas como, a artrite reumatoide e a doença de Crohn. Métodos por cromatografia líquida por exclusão molecular (CL-EM) e por fase reversa (CL-FR) foram desenvolvidos e validados para quantificação de CZP em produtos biotecnológicos. O método por CL-EM foi desenvolvido em coluna cromatográfica BioSep-SEC-S3000 (300 x 4,6 mm, 5μm, 290 Å) mantida à 35°C. A fase móvel A consistiu de 100mM de fosfato de sódio monobásico e 200mM de cloreto de sódio pH 7,0 e a fase móvel B consistiu de Etanol (95:5, v/v), com fluxo de 0,5mL/min e detecção por DAD a 214 nm. No método CL-FR determinou-se a condição analítica com coluna Zorbax 300SB C18 (4.6 x 150 mm, 3,5 µm), mantida a 80°C, com fase móvel A constituída de 0,1% (v/v) de ácido trifluoracético (TFA) em água e fase móvel B elaborada por propanol:acetonitrila:água:TFA (70:20:9,9:0,1, v/v). A eluição ocorreu em gradiente de concentração da fase móvel em fluxo de 1 ml/min e detecção por detector de arranjo de diodos (DAD) em 214nm. O CZP foi eluído nos tempos de retenção de 5,6 e 9,0 min, sendo linear na faixa de concentração de 1 – 40 mg / mL (r² = 0,9993) e 1 – 40 mg / mL (r² = 0,9997), respectivamente, para CLEM e CL–FR. A especificidade dos métodos foi investigada por estudos de degradação forçada, interferência dos excipientes da formulação e análise da pureza dos picos. Os limites de detecção e quantificação foram de 0,14 e 0,41 mg / mL para o método por CL–EM e 0,06 e 0,17 mg / mL para CL–FR. As médias da exatidão foram de 100,50 e 99,80 com bias de 0,85 e 0,82 %, respectivamente, para os métodos por CL-EM e CL–FR. A validação dos métodos seguiu as diretrizes estabelecidas nos guias oficiais da Agência Nacional de Vigilância Sanitária (ANVISA) e da Conferência Internacional de Harmonização (ICH). Além disso, estudou-se bioensaio por cultura celular utilizando a linhagem MONO-MAC-6 estimulada por lipopolissacarídeo (LPS), para avaliação da atividade biológica do CZP e inibição da liberação de TNF-α, que se mostrou significativa. Realizaram-se estudos de correlação entre os métodos validados, visando contribuir para aprimorar o controle de qualidade do produto biotecnológico, no intuito de garantir a segurança e eficácia terapêutica.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESporUniversidade Federal de Santa MariaCentro de Ciências da SaúdePrograma de Pós-Graduação em Ciências FarmacêuticasUFSMBrasilAnálises Clínicas e ToxicológicasAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessCertolizumabe pegolCromatografia líquida por exclusão molecularCromatografia líquida em fase reversaValidaçãoBioensaioCertolizumab pegolSize-exclusion liquid chromatographyReversedphase liquid chromatographyValidationBioassayCNPQ::CIENCIAS DA SAUDE::FARMACIAPesquisa de métodos cromatográficos para quantificação e caracterização do certolizumabe pegolResearch of chromatographic methods for quantification and characterization of certolizumab pegolinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisDalmora, Sergio Luizhttp://lattes.cnpq.br/4505166045049607Silva, Carine VianaMalesuik, Marcelo DonadelSangoi, Maximiliano da SilvaSouza, Fábio Santos dehttp://lattes.cnpq.br/6041086096872362Cardoso Júnior, Clóvis Dervil Appratto400300000005600600600600600600600c5c3d2e7-af0f-4f36-b006-046daf37340a9ea15c85-cc82-4cd1-bf12-28bd1d217ea7c44ed480-6f9b-46d9-b4d3-efeb255f4951549f9437-1bd8-47cc-8e18-e95fd1feeaca9e1df32f-e461-4855-b53e-82250ae7c57405e1bbb9-d6b1-4670-ab86-c8b8c28f8245reponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMLICENSElicense.txtlicense.txttext/plain; charset=utf-81956http://repositorio.ufsm.br/bitstream/1/26640/3/license.txt2f0571ecee68693bd5cd3f17c1e075dfMD53ORIGINALTES_PPGCF_2022_CARDOSO_JÚNIOR_CLÓVIS.pdfTES_PPGCF_2022_CARDOSO_JÚNIOR_CLÓVIS.pdfTese de Doutoradoapplication/pdf1304527http://repositorio.ufsm.br/bitstream/1/26640/1/TES_PPGCF_2022_CARDOSO_J%c3%9aNIOR_CL%c3%93VIS.pdf1aeb3ea1669ab0e1938fe67416649e2eMD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8805http://repositorio.ufsm.br/bitstream/1/26640/2/license_rdf4460e5956bc1d1639be9ae6146a50347MD521/266402022-10-24 11:19:08.782oai:repositorio.ufsm.br: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ório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestopendoar:39132022-10-24T14:19:08Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.por.fl_str_mv |
Pesquisa de métodos cromatográficos para quantificação e caracterização do certolizumabe pegol |
dc.title.alternative.eng.fl_str_mv |
Research of chromatographic methods for quantification and characterization of certolizumab pegol |
title |
Pesquisa de métodos cromatográficos para quantificação e caracterização do certolizumabe pegol |
spellingShingle |
Pesquisa de métodos cromatográficos para quantificação e caracterização do certolizumabe pegol Cardoso Júnior, Clóvis Dervil Appratto Certolizumabe pegol Cromatografia líquida por exclusão molecular Cromatografia líquida em fase reversa Validação Bioensaio Certolizumab pegol Size-exclusion liquid chromatography Reversedphase liquid chromatography Validation Bioassay CNPQ::CIENCIAS DA SAUDE::FARMACIA |
title_short |
Pesquisa de métodos cromatográficos para quantificação e caracterização do certolizumabe pegol |
title_full |
Pesquisa de métodos cromatográficos para quantificação e caracterização do certolizumabe pegol |
title_fullStr |
Pesquisa de métodos cromatográficos para quantificação e caracterização do certolizumabe pegol |
title_full_unstemmed |
Pesquisa de métodos cromatográficos para quantificação e caracterização do certolizumabe pegol |
title_sort |
Pesquisa de métodos cromatográficos para quantificação e caracterização do certolizumabe pegol |
author |
Cardoso Júnior, Clóvis Dervil Appratto |
author_facet |
Cardoso Júnior, Clóvis Dervil Appratto |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Dalmora, Sergio Luiz |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/4505166045049607 |
dc.contributor.referee1.fl_str_mv |
Silva, Carine Viana |
dc.contributor.referee2.fl_str_mv |
Malesuik, Marcelo Donadel |
dc.contributor.referee3.fl_str_mv |
Sangoi, Maximiliano da Silva |
dc.contributor.referee4.fl_str_mv |
Souza, Fábio Santos de |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/6041086096872362 |
dc.contributor.author.fl_str_mv |
Cardoso Júnior, Clóvis Dervil Appratto |
contributor_str_mv |
Dalmora, Sergio Luiz Silva, Carine Viana Malesuik, Marcelo Donadel Sangoi, Maximiliano da Silva Souza, Fábio Santos de |
dc.subject.por.fl_str_mv |
Certolizumabe pegol Cromatografia líquida por exclusão molecular Cromatografia líquida em fase reversa Validação Bioensaio |
topic |
Certolizumabe pegol Cromatografia líquida por exclusão molecular Cromatografia líquida em fase reversa Validação Bioensaio Certolizumab pegol Size-exclusion liquid chromatography Reversedphase liquid chromatography Validation Bioassay CNPQ::CIENCIAS DA SAUDE::FARMACIA |
dc.subject.eng.fl_str_mv |
Certolizumab pegol Size-exclusion liquid chromatography Reversedphase liquid chromatography Validation Bioassay |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS DA SAUDE::FARMACIA |
description |
Certolizumab pegol (CZP) is a fragment of a recombinant humanized monoclonal antibody (mAb) produced in E. coli, conjugated with polyethylene glycol (PEG) that acts by inhibiting the activity of Tumor Necrosis Factor alpha (TNF-α). CZP has a therapeutic indication for the treatment of chronic inflammatory diseases such as rheumatoid arthritis and Crohn's disease. Size exclusion chromatography (SEC) and Reversed phase liquid chromatography (RP-LC) methods were developed and validated for quantification of CZP in biotechnological products. The CL-EM method was developed on a BioSep-SEC-S3000 chromatographic column (300 x 4.6 mm, 5μm, 290 Å) maintained at 35°C. Mobile phase A consisted of 100mM monobasic sodium phosphate and 200mM sodium chloride pH 7.0 and mobile phase B consisted of Ethanol (95:5, v/v), with a flow of 0.5mL/min and detection by DAD at 214 nm. In the CL-FR method, the analytical condition was determined with a Zorbax 300SB C18 column (4.6 x 150 mm, 3.5 µm), maintained at 80°C, with mobile phase A consisting of 0.1% (v/v) of trifluoroacetic acid (TFA) in water and mobile phase B prepared by propanol:acetonitrile:water:TFA (70:20:9,9:0.1, v/v). Elution took place in a mobile phase concentration gradient at a flow of 1 ml/min and detection by a diode array detector (DAD) at 214nm. CZP was eluted at retention times of 5.6 and 9.0 min, being linear in the concentration range of 1 - 40 mg / mL (r² = 0.9993) and 1 - 40 mg / mL (r² = 0.9997), respectively, for SEC and for RP-LC. The specificity of the methods was investigated by forced degradation studies, interference of formulation excipients and analysis of peak purity. The limits of detection and quantification were 0.14 and 0.41 mg / mL for the SEC method and 0.06 and 0.17 mg / mL for RP-LC. The accuracy means were 100.50 and 99.80 with a bias of 0.85 and 0.82%, respectively, for the SEC and RP-LC methods. The validation of the methods followed the guidelines established in the official guides of the National Health Surveillance Agency (ANVISA) and the International Conference on Harmonization (ICH). In addition, a cell culture bioassay using the MONO-MAC-6 stimulated by lipopolysaccharide (LPS) was studied to evaluate the biological activity of CZP and inhibition of TNF-α release, which proved to be significant. Correlation studies were carried out between the validated methods, aiming to contribute to improve the quality control of the biotechnological product, in order to ensure safety and therapeutic efficacy. |
publishDate |
2022 |
dc.date.accessioned.fl_str_mv |
2022-10-24T14:19:08Z |
dc.date.available.fl_str_mv |
2022-10-24T14:19:08Z |
dc.date.issued.fl_str_mv |
2022-05-24 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/26640 |
url |
http://repositorio.ufsm.br/handle/1/26640 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.cnpq.fl_str_mv |
400300000005 |
dc.relation.confidence.fl_str_mv |
600 600 600 600 600 600 600 |
dc.relation.authority.fl_str_mv |
c5c3d2e7-af0f-4f36-b006-046daf37340a 9ea15c85-cc82-4cd1-bf12-28bd1d217ea7 c44ed480-6f9b-46d9-b4d3-efeb255f4951 549f9437-1bd8-47cc-8e18-e95fd1feeaca 9e1df32f-e461-4855-b53e-82250ae7c574 05e1bbb9-d6b1-4670-ab86-c8b8c28f8245 |
dc.rights.driver.fl_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências da Saúde |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Ciências Farmacêuticas |
dc.publisher.initials.fl_str_mv |
UFSM |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
Análises Clínicas e Toxicológicas |
publisher.none.fl_str_mv |
Universidade Federal de Santa Maria Centro de Ciências da Saúde |
dc.source.none.fl_str_mv |
reponame:Manancial - Repositório Digital da UFSM instname:Universidade Federal de Santa Maria (UFSM) instacron:UFSM |
instname_str |
Universidade Federal de Santa Maria (UFSM) |
instacron_str |
UFSM |
institution |
UFSM |
reponame_str |
Manancial - Repositório Digital da UFSM |
collection |
Manancial - Repositório Digital da UFSM |
bitstream.url.fl_str_mv |
http://repositorio.ufsm.br/bitstream/1/26640/3/license.txt http://repositorio.ufsm.br/bitstream/1/26640/1/TES_PPGCF_2022_CARDOSO_J%c3%9aNIOR_CL%c3%93VIS.pdf http://repositorio.ufsm.br/bitstream/1/26640/2/license_rdf |
bitstream.checksum.fl_str_mv |
2f0571ecee68693bd5cd3f17c1e075df 1aeb3ea1669ab0e1938fe67416649e2e 4460e5956bc1d1639be9ae6146a50347 |
bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 |
repository.name.fl_str_mv |
Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM) |
repository.mail.fl_str_mv |
|
_version_ |
1801223802136297472 |