Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1089/mab.2017.0055 http://hdl.handle.net/11449/179831 |
Resumo: | We report an immunization technique that can update the production of monoclonal antibodies (mAbs): the multiple tolerization subtractive immunization (MTSI). A total of 10 BALB/C mice were used. Animals in group 1 received one inoculation of RWPE-1 cells (nontumoral), followed by cyclophosphamide, and then received serial inoculations of nonirradiated PC3 cells (tumoral). Animals in group 2 received our MTSI protocol, as follows: one inoculation of RWPE-1 cells, followed by cyclophosphamide (Cy). This whole tolerization step was repeated three other times, with 14-day intervals between the last Cy exposure and the next RWPE-1 cell inoculation. Finally, the animals received the same nonirradiated PC3 cell exposure as group 1. Blood was taken from each animal, and their polyclonal sera individually tested against the nontumoral RWPE-1 cells in flow cytometry. We found out that, after the MTSI was employed, the serum of the immunized animals, in group 2, contained considerably less antibodies that reacted against the tolerogenic cells, compared with the serum of the animals that underwent regular subtractive immunization. We showed that, by repeating the tolerization cycles, the polyclonal antibodies produced by mice have a reduced specificity toward common/immunodominant epitopes present at nontumoral cells, and thus this technique can be readily used by others in studies involving murine mAb protocols. |
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Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal AntibodiesHybridomaImmunizationMonoclonal antibodiesMurineSubtractive immunizationTolerizationWe report an immunization technique that can update the production of monoclonal antibodies (mAbs): the multiple tolerization subtractive immunization (MTSI). A total of 10 BALB/C mice were used. Animals in group 1 received one inoculation of RWPE-1 cells (nontumoral), followed by cyclophosphamide, and then received serial inoculations of nonirradiated PC3 cells (tumoral). Animals in group 2 received our MTSI protocol, as follows: one inoculation of RWPE-1 cells, followed by cyclophosphamide (Cy). This whole tolerization step was repeated three other times, with 14-day intervals between the last Cy exposure and the next RWPE-1 cell inoculation. Finally, the animals received the same nonirradiated PC3 cell exposure as group 1. Blood was taken from each animal, and their polyclonal sera individually tested against the nontumoral RWPE-1 cells in flow cytometry. We found out that, after the MTSI was employed, the serum of the immunized animals, in group 2, contained considerably less antibodies that reacted against the tolerogenic cells, compared with the serum of the animals that underwent regular subtractive immunization. We showed that, by repeating the tolerization cycles, the polyclonal antibodies produced by mice have a reduced specificity toward common/immunodominant epitopes present at nontumoral cells, and thus this technique can be readily used by others in studies involving murine mAb protocols.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Monoclonal Antibody Laboratory Proteomics Center Univ Estadual Paulista - UNESP School of Pharmaceutical SciencesCell Engineering Laboratory Blood Transfusion Center Univ Estadual Paulista - UNESP School of MedicineDepartment of Bioprocess and Biotechnology Univ Estadual Paulista - UNESP School of Pharmaceutical SciencesMonoclonal Antibody Laboratory Proteomics Center Univ Estadual Paulista - UNESP School of Pharmaceutical SciencesCell Engineering Laboratory Blood Transfusion Center Univ Estadual Paulista - UNESP School of MedicineDepartment of Bioprocess and Biotechnology Univ Estadual Paulista - UNESP School of Pharmaceutical SciencesCNPq: 425535/2016-9Universidade Estadual Paulista (Unesp)De Almeida, Rodrigo [UNESP]Nakamura, Cecília Naomi [UNESP]De Lima Fontes, Marina [UNESP]Da Silva, Julia Paiola [UNESP]Bertanha, Matheus [UNESP]Deffune, Elenice [UNESP]Fusco-Almeida, Ana Marisa [UNESP]Moroz, Andrei [UNESP]2018-12-11T17:36:56Z2018-12-11T17:36:56Z2018-04-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article100-104application/pdfhttp://dx.doi.org/10.1089/mab.2017.0055Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, v. 37, n. 2, p. 100-104, 2018.2167-9436http://hdl.handle.net/11449/17983110.1089/mab.2017.00552-s2.0-850462897672-s2.0-85046289767.pdf4513014379461383Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMonoclonal Antibodies in Immunodiagnosis and Immunotherapy0,421info:eu-repo/semantics/openAccess2023-11-30T06:21:21Zoai:repositorio.unesp.br:11449/179831Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462023-11-30T06:21:21Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies |
title |
Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies |
spellingShingle |
Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies De Almeida, Rodrigo [UNESP] Hybridoma Immunization Monoclonal antibodies Murine Subtractive immunization Tolerization |
title_short |
Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies |
title_full |
Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies |
title_fullStr |
Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies |
title_full_unstemmed |
Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies |
title_sort |
Multiple Tolerization Reduces Antibody Binding Against Tolerogen Cells: Implications for the Production of Monoclonal Antibodies |
author |
De Almeida, Rodrigo [UNESP] |
author_facet |
De Almeida, Rodrigo [UNESP] Nakamura, Cecília Naomi [UNESP] De Lima Fontes, Marina [UNESP] Da Silva, Julia Paiola [UNESP] Bertanha, Matheus [UNESP] Deffune, Elenice [UNESP] Fusco-Almeida, Ana Marisa [UNESP] Moroz, Andrei [UNESP] |
author_role |
author |
author2 |
Nakamura, Cecília Naomi [UNESP] De Lima Fontes, Marina [UNESP] Da Silva, Julia Paiola [UNESP] Bertanha, Matheus [UNESP] Deffune, Elenice [UNESP] Fusco-Almeida, Ana Marisa [UNESP] Moroz, Andrei [UNESP] |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
De Almeida, Rodrigo [UNESP] Nakamura, Cecília Naomi [UNESP] De Lima Fontes, Marina [UNESP] Da Silva, Julia Paiola [UNESP] Bertanha, Matheus [UNESP] Deffune, Elenice [UNESP] Fusco-Almeida, Ana Marisa [UNESP] Moroz, Andrei [UNESP] |
dc.subject.por.fl_str_mv |
Hybridoma Immunization Monoclonal antibodies Murine Subtractive immunization Tolerization |
topic |
Hybridoma Immunization Monoclonal antibodies Murine Subtractive immunization Tolerization |
description |
We report an immunization technique that can update the production of monoclonal antibodies (mAbs): the multiple tolerization subtractive immunization (MTSI). A total of 10 BALB/C mice were used. Animals in group 1 received one inoculation of RWPE-1 cells (nontumoral), followed by cyclophosphamide, and then received serial inoculations of nonirradiated PC3 cells (tumoral). Animals in group 2 received our MTSI protocol, as follows: one inoculation of RWPE-1 cells, followed by cyclophosphamide (Cy). This whole tolerization step was repeated three other times, with 14-day intervals between the last Cy exposure and the next RWPE-1 cell inoculation. Finally, the animals received the same nonirradiated PC3 cell exposure as group 1. Blood was taken from each animal, and their polyclonal sera individually tested against the nontumoral RWPE-1 cells in flow cytometry. We found out that, after the MTSI was employed, the serum of the immunized animals, in group 2, contained considerably less antibodies that reacted against the tolerogenic cells, compared with the serum of the animals that underwent regular subtractive immunization. We showed that, by repeating the tolerization cycles, the polyclonal antibodies produced by mice have a reduced specificity toward common/immunodominant epitopes present at nontumoral cells, and thus this technique can be readily used by others in studies involving murine mAb protocols. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-12-11T17:36:56Z 2018-12-11T17:36:56Z 2018-04-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1089/mab.2017.0055 Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, v. 37, n. 2, p. 100-104, 2018. 2167-9436 http://hdl.handle.net/11449/179831 10.1089/mab.2017.0055 2-s2.0-85046289767 2-s2.0-85046289767.pdf 4513014379461383 |
url |
http://dx.doi.org/10.1089/mab.2017.0055 http://hdl.handle.net/11449/179831 |
identifier_str_mv |
Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, v. 37, n. 2, p. 100-104, 2018. 2167-9436 10.1089/mab.2017.0055 2-s2.0-85046289767 2-s2.0-85046289767.pdf 4513014379461383 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Monoclonal Antibodies in Immunodiagnosis and Immunotherapy 0,421 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
100-104 application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1797789866937810944 |