Morphological selection of parental Chinese hamster ovary cell clones exhibiting high-level expression of recombinant protein

Citation
Jy. Chung et al., Morphological selection of parental Chinese hamster ovary cell clones exhibiting high-level expression of recombinant protein, BIOTECHNIQU, 29(4), 2000, pp. 768
Citations number
13
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOTECHNIQUES
ISSN journal
07366205 → ACNP
Volume
29
Issue
4
Year of publication
2000
Database
ISI
SICI code
0736-6205(200010)29:4<768:MSOPCH>2.0.ZU;2-8
Abstract
To facilitate the establishment of recombinant Chinese hamster ovary (rCHO) cell lines with dihydrofolate reductase (dhfr)mediated gene amplification, a primary selection method based on morphology of parental CHO clones has been developed Morphology of parental clones that were made by transfecting various plasmids encoding thrombopoietin (TPO) and its analogs and humaniz ed antibodies into dhfr-deficient (dhfr(-)) CHO cells was not uniform. Morp hology of many parental clones exhibiting high-level expression of the intr oduced gene was similar to that of nontransfected dhfr- CHO cells. On the o ther hand, most parental clones with low-level expression experienced notic eable morphological changes such as bipolar fibroblast-like morphology. In case of selection of parental clones with TPO expression level higher than 200 ng/mL, morphological selection improved selection efficiency by 3.5-fol d compared with random selection. Furthermore, when subjected to methotrexa te for gene amplification, parental clones that were selected based on morp hology elevated the expression level as much as those that were selected ra ndomly. Taken together morphological selection of parental clones can facil itate the establishment of rCHO cell lines expressing recombinant proteins.