Development of a Mammalian Cell Line for Stable Production of Anti-PD-1
<b>Background/Objectives:</b> Immune checkpoint blockade, particularly targeting the programmed cell death 1 (PD-1) receptor, is a promising strategy in cancer immunotherapy. The interaction between PD-1 and its ligands, PD-L1 and PD-L2, is crucial in immune evasion by tumors. Blocking t...
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MDPI AG
2024-10-01
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| author | Erika Csató-Kovács Pál Salamon Szilvia Fikó-Lászlo Krisztina Kovács Alice Koka Mónika András-Korodi Emőke Antal Emília Brumă Brigitta Tőrsők Szilárd Gudor Ildikó Miklóssy Kálmán Csongor Orbán Csilla Albert Emese Éva Bálint Beáta Albert |
| author_facet | Erika Csató-Kovács Pál Salamon Szilvia Fikó-Lászlo Krisztina Kovács Alice Koka Mónika András-Korodi Emőke Antal Emília Brumă Brigitta Tőrsők Szilárd Gudor Ildikó Miklóssy Kálmán Csongor Orbán Csilla Albert Emese Éva Bálint Beáta Albert |
| author_sort | Erika Csató-Kovács |
| collection | DOAJ |
| description | <b>Background/Objectives:</b> Immune checkpoint blockade, particularly targeting the programmed cell death 1 (PD-1) receptor, is a promising strategy in cancer immunotherapy. The interaction between PD-1 and its ligands, PD-L1 and PD-L2, is crucial in immune evasion by tumors. Blocking this interaction with monoclonal antibodies like Nivolumab can restore anti-tumor immunity. This study aims to develop a stable expression system for Nivolumab-based anti-PD-1 in the Chinese Hamster Ovary (CHO) DG44 cell line using two different expression vector systems with various signal sequences. <b>Methods:</b> The heavy chain (HC) and light chain (LC) of Nivolumab were cloned into two expression vectors, pOptiVEC and pcDNA3.3. Each vector was engineered with two distinct signal sequences, resulting in the creation of eight recombinant plasmids. These plasmids were co-transfected into CHO DG44 cells in different combinations, allowing for the assessment of stable antibody production. <b>Results:</b> Both pOptiVEC and pcDNA3.3 vectors were successful in stably integrating and expressing the Nivolumab-based anti-PD-1 antibody in CHO DG44 cells. This study found that the choice of signal sequence significantly influenced the quantity of antibodies produced. The optimization of production conditions further enhanced antibody yield, indicating the potential for large-scale production. <b>Conclusions:</b> This study demonstrates that both pOptiVEC and pcDNA3.3 expression systems are effective for the stable production of Nivolumab-based anti-PD-1 in CHO DG44 cells. Signal sequences play a critical role in determining the expression levels, and optimizing production conditions can further increase antibody yield, supporting future applications in cancer immunotherapy. |
| format | Article |
| id | doaj-art-70c1a9aa0dde46af8ee25d8e6185e1eb |
| institution | OA Journals |
| issn | 2073-4468 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibodies |
| spelling | doaj-art-70c1a9aa0dde46af8ee25d8e6185e1eb2025-08-20T02:00:59ZengMDPI AGAntibodies2073-44682024-10-011348210.3390/antib13040082Development of a Mammalian Cell Line for Stable Production of Anti-PD-1Erika Csató-Kovács0Pál Salamon1Szilvia Fikó-Lászlo2Krisztina Kovács3Alice Koka4Mónika András-Korodi5Emőke Antal6Emília Brumă7Brigitta Tőrsők8Szilárd Gudor9Ildikó Miklóssy10Kálmán Csongor Orbán11Csilla Albert12Emese Éva Bálint13Beáta Albert14Department of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, RomaniaDepartment of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, RomaniaFaculty of Natural Sciences, University of Pécs, 7 Ifjúság Útja St., 7624 Pécs, HungaryFaculty of Natural Sciences, University of Pécs, 7 Ifjúság Útja St., 7624 Pécs, HungaryDepartment of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, RomaniaCorax-Bioner CEU S.A., 1 Miko Str., et. 1, Cam. 100, 530174 Miercurea Ciuc, RomaniaCorax-Bioner CEU S.A., 1 Miko Str., et. 1, Cam. 100, 530174 Miercurea Ciuc, RomaniaFaculty of Natural Sciences, University of Pécs, 7 Ifjúság Útja St., 7624 Pécs, HungaryFaculty of Natural Sciences, University of Pécs, 7 Ifjúság Útja St., 7624 Pécs, HungaryDepartment of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, RomaniaDepartment of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, RomaniaDepartment of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, RomaniaDepartment of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, RomaniaDepartment of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, RomaniaDepartment of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, Romania<b>Background/Objectives:</b> Immune checkpoint blockade, particularly targeting the programmed cell death 1 (PD-1) receptor, is a promising strategy in cancer immunotherapy. The interaction between PD-1 and its ligands, PD-L1 and PD-L2, is crucial in immune evasion by tumors. Blocking this interaction with monoclonal antibodies like Nivolumab can restore anti-tumor immunity. This study aims to develop a stable expression system for Nivolumab-based anti-PD-1 in the Chinese Hamster Ovary (CHO) DG44 cell line using two different expression vector systems with various signal sequences. <b>Methods:</b> The heavy chain (HC) and light chain (LC) of Nivolumab were cloned into two expression vectors, pOptiVEC and pcDNA3.3. Each vector was engineered with two distinct signal sequences, resulting in the creation of eight recombinant plasmids. These plasmids were co-transfected into CHO DG44 cells in different combinations, allowing for the assessment of stable antibody production. <b>Results:</b> Both pOptiVEC and pcDNA3.3 vectors were successful in stably integrating and expressing the Nivolumab-based anti-PD-1 antibody in CHO DG44 cells. This study found that the choice of signal sequence significantly influenced the quantity of antibodies produced. The optimization of production conditions further enhanced antibody yield, indicating the potential for large-scale production. <b>Conclusions:</b> This study demonstrates that both pOptiVEC and pcDNA3.3 expression systems are effective for the stable production of Nivolumab-based anti-PD-1 in CHO DG44 cells. Signal sequences play a critical role in determining the expression levels, and optimizing production conditions can further increase antibody yield, supporting future applications in cancer immunotherapy.https://www.mdpi.com/2073-4468/13/4/82immune checkpoint moleculeanti-PD-1CHO DG44Nivolumabmonoclonal antibody production |
| spellingShingle | Erika Csató-Kovács Pál Salamon Szilvia Fikó-Lászlo Krisztina Kovács Alice Koka Mónika András-Korodi Emőke Antal Emília Brumă Brigitta Tőrsők Szilárd Gudor Ildikó Miklóssy Kálmán Csongor Orbán Csilla Albert Emese Éva Bálint Beáta Albert Development of a Mammalian Cell Line for Stable Production of Anti-PD-1 Antibodies immune checkpoint molecule anti-PD-1 CHO DG44 Nivolumab monoclonal antibody production |
| title | Development of a Mammalian Cell Line for Stable Production of Anti-PD-1 |
| title_full | Development of a Mammalian Cell Line for Stable Production of Anti-PD-1 |
| title_fullStr | Development of a Mammalian Cell Line for Stable Production of Anti-PD-1 |
| title_full_unstemmed | Development of a Mammalian Cell Line for Stable Production of Anti-PD-1 |
| title_short | Development of a Mammalian Cell Line for Stable Production of Anti-PD-1 |
| title_sort | development of a mammalian cell line for stable production of anti pd 1 |
| topic | immune checkpoint molecule anti-PD-1 CHO DG44 Nivolumab monoclonal antibody production |
| url | https://www.mdpi.com/2073-4468/13/4/82 |
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