title: Expression of chimeric antibody in mammalian cells using dicistronic expression vector.
abstract: 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.
authors.affiliation: Department of Anatomy, Fourth Military Medical University, 710032, Xi'an City, China. liangjh@fmmu.edu.cn
authors.forename: Kang-Hui
authors.initials: KH
authors.lastname: Xiong
authors.forename: Qin-Chuan
authors.initials: QC
authors.lastname: Liang
authors.forename: Hua
authors.initials: H
authors.lastname: Xiong
authors.forename: Chang-Xu
authors.initials: CX
authors.lastname: Zou
authors.forename: Guo-Dong
authors.initials: GD
authors.lastname: Gao
authors.forename: Zhen-Wei
authors.initials: ZW
authors.lastname: Zhao
authors.forename: Hua
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D016466:CHO Cells; D006224:Cricetinae; D015870:Gene Expression; D005796:Genes; D005822:Genetic Vectors; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D013762:Tetrahydrofolate Dehydrogenase / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D013762:Tetrahydrofolate Dehydrogenase