Evaluating the expression profile and stability of different UCOE containing vector combinations in mAb-producing CHO cells
| Autor: | Samira Ahmadi, Fatemeh Davami, Vahid Khalaj, Fatemeh Nematpour, Fereidoun Mahboudi, Behrouz Vaziri, Maryam Ahmadi, Saedeh Ebadat |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Cell line development
0106 biological sciences 0301 basic medicine medicine.drug_class Genetic Vectors Cell CHO Cells Biology Protein Engineering Immunoglobulin light chain Monoclonal antibody 01 natural sciences Monoclonal antibody (mAb) Chinese hamster ovary (CHO) 03 medical and health sciences Cricetulus 010608 biotechnology medicine Animals Transgenes Ubiquitous chromatin opening elements (UCOE) Promoter Regions Genetic Gene Chinese hamster ovary cell Antibodies Monoclonal Molecular biology Chromatin 030104 developmental biology medicine.anatomical_structure Gene Expression Regulation Cell culture biology.protein Antibody Research Article Biotechnology |
| Zdroj: | BMC Biotechnology |
| ISSN: | 1472-6750 |
| DOI: | 10.1186/s12896-017-0330-0 |
| Popis: | Background As the demand for monoclonal antibodies (mAb) increases, more efficient expression methods are required for their manufacturing process. Transcriptional gene silencing is a common phenomenon in recombinant cell lines which leads to expression reduction and instability. There are reports on improved antibody expression in ubiquitous chromatin opening element (UCOE) containing both heavy and light chain gene constructs. Here we investigate the impact of having these elements as part of the light chain, heavy chain or both genes during cell line development. In this regard, non-UCOE and UCOE vectors were constructed and stable Chinese hamster ovary (CHO) cell pools were generated by different vector combinations. Results Expression analysis revealed that all UCOE cell pools had higher antibody yields compared to non-UCOE cells, Moreover the most optimal expression was obtained by cells containing just the UCOE on heavy chain. In terms of stability, it was shown that the high level of expression was kept consistence for more than four months in these cells whereas the expression titers were reduced in the other UCOE pools. Conclusions In conclusion, UCOE significantly enhanced the level and stability of antibody expression and the use of this element with heavy chain provided more stable cell lines with higher production level. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|