{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,10]],"date-time":"2026-01-10T20:45:25Z","timestamp":1768077925589,"version":"3.49.0"},"reference-count":26,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2024,10,3]],"date-time":"2024-10-03T00:00:00Z","timestamp":1727913600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Research, Innovation and Digitalization","award":["POC\/161\/6\/3-398\/390074\/09.11.2021"],"award-info":[{"award-number":["POC\/161\/6\/3-398\/390074\/09.11.2021"]}]},{"name":"Collegium Talentum Programme of Hungary","award":["POC\/161\/6\/3-398\/390074\/09.11.2021"],"award-info":[{"award-number":["POC\/161\/6\/3-398\/390074\/09.11.2021"]}]},{"name":"\u00daNKP-23-4-II New National Excellence Program of the Ministry for Innovation and Technology","award":["POC\/161\/6\/3-398\/390074\/09.11.2021"],"award-info":[{"award-number":["POC\/161\/6\/3-398\/390074\/09.11.2021"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibodies"],"abstract":"<jats:p>Background\/Objectives: 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. Methods: 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. Results: 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. Conclusions: 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.<\/jats:p>","DOI":"10.3390\/antib13040082","type":"journal-article","created":{"date-parts":[[2024,10,3]],"date-time":"2024-10-03T03:36:03Z","timestamp":1727926563000},"page":"82","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Development of a Mammalian Cell Line for Stable Production of Anti-PD-1"],"prefix":"10.3390","volume":"13","author":[{"given":"Erika","family":"Csat\u00f3-Kov\u00e1cs","sequence":"first","affiliation":[{"name":"Department of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, Romania"},{"name":"Corax-Bioner CEU S.A., 1 Miko Str., et. 1, Cam. 100, 530174 Miercurea Ciuc, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6803-1212","authenticated-orcid":false,"given":"P\u00e1l","family":"Salamon","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, Romania"},{"name":"Corax-Bioner CEU S.A., 1 Miko Str., et. 1, Cam. 100, 530174 Miercurea Ciuc, Romania"},{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Szilvia","family":"Fik\u00f3-L\u00e1szlo","sequence":"additional","affiliation":[{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Krisztina","family":"Kov\u00e1cs","sequence":"additional","affiliation":[{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alice","family":"Koka","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M\u00f3nika","family":"Andr\u00e1s-Korodi","sequence":"additional","affiliation":[{"name":"Corax-Bioner CEU S.A., 1 Miko Str., et. 1, Cam. 100, 530174 Miercurea Ciuc, Romania"},{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Em\u0151ke","family":"Antal","sequence":"additional","affiliation":[{"name":"Corax-Bioner CEU S.A., 1 Miko Str., et. 1, Cam. 100, 530174 Miercurea Ciuc, Romania"},{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Em\u00edlia","family":"Brum\u0103","sequence":"additional","affiliation":[{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brigitta","family":"T\u0151rs\u0151k","sequence":"additional","affiliation":[{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Szil\u00e1rd","family":"Gudor","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8037-7216","authenticated-orcid":false,"given":"Ildik\u00f3","family":"Mikl\u00f3ssy","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, Romania"},{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"},{"name":"C. 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Neni\u021bescu Institute of Organic Chemistry, 202B Splaiul Independen\u021bei, Sector 6, 060023 Bucure\u0219ti, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7509-748X","authenticated-orcid":false,"given":"K\u00e1lm\u00e1n Csongor","family":"Orb\u00e1n","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, Romania"},{"name":"Corax-Bioner CEU S.A., 1 Miko Str., et. 1, Cam. 100, 530174 Miercurea Ciuc, Romania"},{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6882-6691","authenticated-orcid":false,"given":"Csilla","family":"Albert","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, Romania"},{"name":"Corax-Bioner CEU S.A., 1 Miko Str., et. 1, Cam. 100, 530174 Miercurea Ciuc, Romania"},{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emese \u00c9va","family":"B\u00e1lint","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, Romania"},{"name":"Corax-Bioner CEU S.A., 1 Miko Str., et. 1, Cam. 100, 530174 Miercurea Ciuc, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Be\u00e1ta","family":"Albert","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Faculty of Economics, Socio-Human Sciences and Engineering, Sapientia Hungarian University of Transylvania, 1 Libertatii Sq, 530104 Miercurea Ciuc, Romania"},{"name":"Corax-Bioner CEU S.A., 1 Miko Str., et. 1, Cam. 100, 530174 Miercurea Ciuc, Romania"},{"name":"Faculty of Natural Sciences, University of P\u00e9cs, 7 Ifj\u00fas\u00e1g \u00datja St., 7624 P\u00e9cs, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"466","DOI":"10.4161\/mabs.2.5.12720","article-title":"Cell Culture Processes for Monoclonal Antibody Production","volume":"2","author":"Li","year":"2010","journal-title":"MAbs"},{"key":"ref_2","unstructured":"Barentine, C. 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