Expression of chimeric antibody in mammalian cells using dicistronic expression vector

• Published in: Biotechnology letters Vol. 27; no. 21; pp. 1713 - 1717
• Main Authors: XIONG, Kang-Hui, LIANG, Qin-Chuan, HUA XIONG, ZOU, Chang-Xu, GAO, Guo-Dong, ZHAO, Zhen-Wei, HUA ZHANG
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.11.2005; Springer Nature B.V
• Subjects: Animals; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - genetics; Biological and medical sciences; Biotechnology; CHO Cells; Cricetinae; Fundamental and applied biological sciences. Psychology; Gene Expression; Genes; Genetic Vectors; Proteins; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - genetics; Rodents; Tetrahydrofolate Dehydrogenase - biosynthesis; Tetrahydrofolate Dehydrogenase - genetics; chimeric antibody; Enzyme; Antibody; Gene expression; CHO cell line; Chinese hamster ovary cells; antibody engineering; Vertebrata; Mammalia; Oxidoreductases; Dihydrofolate reductase; dicistronic expression vector; Vector
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-005-2736-3

A dicistronic expression vector was constructed for Chinese hamster ovary (CHO) cells that produce both selectable marker-DHFR (dihydrofolate reductase) gene and recombinant antibody cDNA from a single primary transcript via differential splicing. The vector was derived from a pDHL vector and contained the human constant region cDNA so that any human-mouse chimeric antibodies could be expressed. The expression vector produced stable CHO cell clones that secreted nearly double the amount of chimeric antibodies than produced by conventional expression approaches, where the DHFR gene and relevant cDNA are controlled by separate transcription cassettes. Clones with increased expression of interested genes can be efficiently generated by selection in medium containing a gradually increasing amount of methotrexate. The dicistronic expression system using incomplete splicing DHFR gene strategy thus provides a convenient, high-level, and rapid expression of chimeric antibodies.