Chapter title |
An IRES-Mediated Tricistronic Vector for Efficient Generation of Stable, High-Level Monoclonal Antibody Producing CHO DG44 Cell Lines
|
---|---|
Chapter number | 17 |
Book title |
Antibody Engineering
|
Published in |
Methods in molecular biology, September 2018
|
DOI | 10.1007/978-1-4939-8648-4_17 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8647-7, 978-1-4939-8648-4
|
Authors |
Jessna H. M. Yeo, Mariati, Yuansheng Yang, Yeo, Jessna H. M., Mariati, , Yang, Yuansheng |
Abstract |
The generation of stable, high-level monoclonal antibody (mAb) producing cell lines remains a major challenge in biopharmaceutical industry. The commonly used plasmid vectors for mAb expression, which express light chain (LC), heavy chain (HC), and selection marker genes on separate vectors or via multiple promoters on a single vector, are not able to accurately control the ratio of LC over HC expression and tend to result in non-expressing clones. To overcome these issues, we have developed a tricistronic vector using two internal ribosome entry sites (IRES) to express the LC, HC, and dihydrofolate reductase (DHFR) selection marker genes in one transcript. In this tricistronic vector, the three genes are under the control of a hapten-modified human cytomegalovirus (hCMV) promoter containing a core CpG island element (IE) to enhance the production stability. The LC gene is arranged as the first cistron followed by a wild-type IRES to control the HC expression. Such design expresses excess LC polypeptides which enhance mAb expression level and reduce aggregate. A mutated IRES with attenuated strength is applied on DHFR to reduce its expression for enhancing the stringency of selection for high producers. This vector allows easy generation of stable, high mAb producing CHO DG44 pools and clones for antibody development and manufacturing. |
X Demographics
As of 1 July 2024, you may notice a temporary increase in the numbers of X profiles with Unknown location. Click here to learn more.
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 24 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 4 | 17% |
Other | 3 | 13% |
Student > Doctoral Student | 3 | 13% |
Student > Ph. D. Student | 3 | 13% |
Student > Bachelor | 1 | 4% |
Other | 2 | 8% |
Unknown | 8 | 33% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 12 | 50% |
Chemical Engineering | 1 | 4% |
Agricultural and Biological Sciences | 1 | 4% |
Unknown | 10 | 42% |