Publicação
Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody
| Resumo: | Monoclonal antibody-based therapy has shown efficacy against cancer, autoimmune, infectious, and inflammatory diseases. Multispecific antibodies (MsAbs), including trispecifics (tsAbs), offer enhanced therapeutic potential by targeting different epitopes. However, when co-expressed from three or more different polypeptide chains, MsAb production can lead to incorrect chain assembly and co-production of mispaired species with impaired biological activity. Moreover, mispairing carries significant challenges for downstream purification, decreasing yields and increasing the cost of bioprocess development. In this study, quantitative transcriptomics and proteomics analyses were employed to investigate which signaling pathways correlated with low and high mispairing clone signatures. Gene and protein expression profiles of Chinese hamster ovary (CHO) clones producing an tsAb were analyzed in the exponential growth and stationary (tsAb production) phase of fed-batch culture. Functional analysis revealed activated endoplasmic reticulum stress in high mispairing clones in both culture phases, while low mispairing clones exhibited expression profiles indicative of activated protein translation, as well as higher endocytosis and target protein degradation, suggesting the clearance of unfolded proteins through ubiquitin-mediated mechanisms. In addition, through transcriptomic profiling, we identified a group of genes that have the potential to be used as a biomarker panel tool for identifying high mispairing levels in the early stages of bioprocess development. |
|---|---|
| Autores principais: | Sebastião, Maria João |
| Outros Autores: | Hoffman, Michael; Escandell, José; Tousi, Fatemeh; Zhang, Jin; Figueroa, Bruno; DeMaria, Christine; Gomes-Alves, Patrícia |
| Assunto: | CHO cells mispairing multispecific antibodies proteomics transcriptomics Medicine (miscellaneous) General Biochemistry,Genetics and Molecular Biology SDG 3 - Good Health and Well-being |
| Ano: | 2023 |
| País: | Portugal |
| Tipo de documento: | artigo |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade Nova de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Institucional da UNL |
| _version_ | 1868983280905224192 |
|---|---|
| author | Sebastião, Maria João |
| author2 | Hoffman, Michael Escandell, José Tousi, Fatemeh Zhang, Jin Figueroa, Bruno DeMaria, Christine Gomes-Alves, Patrícia |
| author2_role | author author author author author author author |
| author_facet | Sebastião, Maria João Hoffman, Michael Escandell, José Tousi, Fatemeh Zhang, Jin Figueroa, Bruno DeMaria, Christine Gomes-Alves, Patrícia |
| author_role | author |
| contributor_name_str_mv | Instituto de Tecnologia Química e Biológica António Xavier (ITQB) MDPI AG RUN |
| country_str | PT |
| creators_json_txt | [{\"Person.name\":\"Sebastião, Maria João\"},{\"Person.name\":\"Hoffman, Michael\"},{\"Person.name\":\"Escandell, José\"},{\"Person.name\":\"Tousi, Fatemeh\"},{\"Person.name\":\"Zhang, Jin\"},{\"Person.name\":\"Figueroa, Bruno\"},{\"Person.name\":\"DeMaria, Christine\"},{\"Person.name\":\"Gomes-Alves, Patrícia\"}] |
| datacite.contributors.contributor.contributorName.fl_str_mv | Instituto de Tecnologia Química e Biológica António Xavier (ITQB) MDPI AG RUN |
| datacite.creators.creator.creatorName.fl_str_mv | Sebastião, Maria João Hoffman, Michael Escandell, José Tousi, Fatemeh Zhang, Jin Figueroa, Bruno DeMaria, Christine Gomes-Alves, Patrícia |
| datacite.date.Accepted.fl_str_mv | 2023-11-01T00:00:00Z |
| datacite.date.available.fl_str_mv | 2024-04-02T23:52:23Z |
| datacite.date.embargoed.fl_str_mv | 2024-04-02T23:52:23Z |
| datacite.rights.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| datacite.subjects.subject.fl_str_mv | CHO cells mispairing multispecific antibodies proteomics transcriptomics Medicine (miscellaneous) General Biochemistry,Genetics and Molecular Biology SDG 3 - Good Health and Well-being |
| datacite.titles.title.fl_str_mv | Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody |
| dc.contributor.none.fl_str_mv | Instituto de Tecnologia Química e Biológica António Xavier (ITQB) MDPI AG RUN |
| dc.creator.none.fl_str_mv | Sebastião, Maria João Hoffman, Michael Escandell, José Tousi, Fatemeh Zhang, Jin Figueroa, Bruno DeMaria, Christine Gomes-Alves, Patrícia |
| dc.date.Accepted.fl_str_mv | 2023-11-01T00:00:00Z |
| dc.date.available.fl_str_mv | 2024-04-02T23:52:23Z |
| dc.date.embargoed.fl_str_mv | 2024-04-02T23:52:23Z |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | http://hdl.handle.net/10362/165743 |
| dc.language.none.fl_str_mv | eng |
| dc.rights.none.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| dc.subject.none.fl_str_mv | CHO cells mispairing multispecific antibodies proteomics transcriptomics Medicine (miscellaneous) General Biochemistry,Genetics and Molecular Biology SDG 3 - Good Health and Well-being |
| dc.title.fl_str_mv | Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody |
| dc.type.none.fl_str_mv | http://purl.org/coar/resource_type/c_6501 |
| description | Monoclonal antibody-based therapy has shown efficacy against cancer, autoimmune, infectious, and inflammatory diseases. Multispecific antibodies (MsAbs), including trispecifics (tsAbs), offer enhanced therapeutic potential by targeting different epitopes. However, when co-expressed from three or more different polypeptide chains, MsAb production can lead to incorrect chain assembly and co-production of mispaired species with impaired biological activity. Moreover, mispairing carries significant challenges for downstream purification, decreasing yields and increasing the cost of bioprocess development. In this study, quantitative transcriptomics and proteomics analyses were employed to investigate which signaling pathways correlated with low and high mispairing clone signatures. Gene and protein expression profiles of Chinese hamster ovary (CHO) clones producing an tsAb were analyzed in the exponential growth and stationary (tsAb production) phase of fed-batch culture. Functional analysis revealed activated endoplasmic reticulum stress in high mispairing clones in both culture phases, while low mispairing clones exhibited expression profiles indicative of activated protein translation, as well as higher endocytosis and target protein degradation, suggesting the clearance of unfolded proteins through ubiquitin-mediated mechanisms. In addition, through transcriptomic profiling, we identified a group of genes that have the potential to be used as a biomarker panel tool for identifying high mispairing levels in the early stages of bioprocess development. |
| dirty | 0 |
| eu_rights_str_mv | openAccess |
| format | article |
| fulltext.url.fl_str_mv | https://run.unl.pt/bitstreams/a892e93f-9ee4-43b0-b9fb-98055396159c/download |
| id | run_ac2bef3cbafc8dc1e44019a5f596fb34 |
| identifier.url.fl_str_mv | http://hdl.handle.net/10362/165743 |
| inst_facet_str | urn:organizationAcronym:unl{{{_:::_}}}Universidade Nova de Lisboa |
| instacron_str | unl |
| institution | Universidade Nova de Lisboa |
| instname_str | Universidade Nova de Lisboa |
| language | eng |
| network_acronym_str | run |
| network_name_str | Repositório Institucional da UNL |
| oai_identifier_str | oai:run.unl.pt:10362/165743 |
| organization_str_mv | urn:organizationAcronym:unl |
| person_str_mv | Sebastião, Maria João Hoffman, Michael Escandell, José Tousi, Fatemeh Zhang, Jin Figueroa, Bruno DeMaria, Christine Gomes-Alves, Patrícia |
| publishDate | 2023 |
| repo_facet_str | urn:repositoryAcronym:run{{{_:::_}}}Repositório Institucional da UNL |
| reponame_str | Repositório Institucional da UNL |
| repository_id_str | urn:repositoryAcronym:run |
| service_str_mv | urn:repositoryAcronym:run |
| spelling | engenMonoclonal antibody-based therapy has shown efficacy against cancer, autoimmune, infectious, and inflammatory diseases. Multispecific antibodies (MsAbs), including trispecifics (tsAbs), offer enhanced therapeutic potential by targeting different epitopes. However, when co-expressed from three or more different polypeptide chains, MsAb production can lead to incorrect chain assembly and co-production of mispaired species with impaired biological activity. Moreover, mispairing carries significant challenges for downstream purification, decreasing yields and increasing the cost of bioprocess development. In this study, quantitative transcriptomics and proteomics analyses were employed to investigate which signaling pathways correlated with low and high mispairing clone signatures. Gene and protein expression profiles of Chinese hamster ovary (CHO) clones producing an tsAb were analyzed in the exponential growth and stationary (tsAb production) phase of fed-batch culture. Functional analysis revealed activated endoplasmic reticulum stress in high mispairing clones in both culture phases, while low mispairing clones exhibited expression profiles indicative of activated protein translation, as well as higher endocytosis and target protein degradation, suggesting the clearance of unfolded proteins through ubiquitin-mediated mechanisms. In addition, through transcriptomic profiling, we identified a group of genes that have the potential to be used as a biomarker panel tool for identifying high mispairing levels in the early stages of bioprocess development.application/pdfenIdentification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific AntibodySebastião, Maria JoãoHoffman, MichaelEscandell, JoséTousi, FatemehZhang, JinFigueroa, BrunoDeMaria, ChristineGomes-Alves, PatríciaInstituto de Tecnologia Química e Biológica António Xavier (ITQB)MDPI AGHostingInstitutionOrganizationalRUNe-mailmailto:run@unl.ptrun@unl.ptISSNIsPartOf2227-9059URNIsPartOfPURE: 83729506URNIsPartOfPURE UUID: 7e7c218b-2d4c-49f1-be10-2f32f14a083cURNIsPartOfScopus: 85178360671DOIIsPartOf10.3390/biomedicines111128902024-04-02T23:52:23Z2023-112023-11-01T00:00:00ZHandlehttp://hdl.handle.net/10362/165743http://purl.org/coar/access_right/c_abf2open accessCHO cellsmispairingmultispecific antibodiesproteomicstranscriptomicsMedicine (miscellaneous)General Biochemistry,Genetics and Molecular BiologySDG 3 - Good Health and Well-being3081595 bytesliteraturehttp://purl.org/coar/resource_type/c_6501journal articlehttp://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://run.unl.pt/bitstreams/a892e93f-9ee4-43b0-b9fb-98055396159c/download |
| spellingShingle | Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody Sebastião, Maria João CHO cells mispairing multispecific antibodies proteomics transcriptomics Medicine (miscellaneous) General Biochemistry,Genetics and Molecular Biology SDG 3 - Good Health and Well-being |
| status | SINGLETON |
| subject.fl_str_mv | CHO cells mispairing multispecific antibodies proteomics transcriptomics Medicine (miscellaneous) General Biochemistry,Genetics and Molecular Biology SDG 3 - Good Health and Well-being |
| title | Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody |
| title_full | Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody |
| title_fullStr | Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody |
| title_full_unstemmed | Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody |
| title_short | Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody |
| title_sort | Identification of Mispairing Omic Signatures in Chinese Hamster Ovary (CHO) Cells Producing a Tri-Specific Antibody |
| topic | CHO cells mispairing multispecific antibodies proteomics transcriptomics Medicine (miscellaneous) General Biochemistry,Genetics and Molecular Biology SDG 3 - Good Health and Well-being |
| topic_facet | CHO cells mispairing multispecific antibodies proteomics transcriptomics Medicine (miscellaneous) General Biochemistry,Genetics and Molecular Biology SDG 3 - Good Health and Well-being |
| url | http://hdl.handle.net/10362/165743 |
| visible | 1 |