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Titolo:
Morphological selection of parental Chinese hamster ovary cell clones exhibiting high-level expression of recombinant protein
Autore:
Chung, JY; Kim, TK; Lee, GM;
Indirizzi:
Korea Adv Inst Sci & Technol, Dept Biol Sci, Yusong Gu, Taejon 305701, South Korea Korea Adv Inst Sci & Technol Taejon South Korea 305701 5701, South Korea
Titolo Testata:
BIOTECHNIQUES
fascicolo: 4, volume: 29, anno: 2000,
pagine: 768 -
SICI:
0736-6205(200010)29:4<768:MSOPCH>2.0.ZU;2-8
Fonte:
ISI
Lingua:
ENG
Soggetto:
CHO CELLS; TRANSFECTED CELLS; STABILITY; PRESSURE; ABSENCE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
13
Recensione:
Indirizzi per estratti:
Indirizzo: Lee, GM Korea Adv Inst Sci & Technol, Dept Biol Sci, Yusong Gu, 373-1 Kusong Dong,Taejon 305701, South Korea Korea Adv Inst Sci & Technol 373-1 Kusong Dong Taejon South Korea 305701
Citazione:
J.Y. 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

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 hasbeen developed Morphology of parental clones that were made by transfecting various plasmids encoding thrombopoietin (TPO) and its analogs and humanized antibodies into dhfr-deficient (dhfr(-)) CHO cells was not uniform. Morphology of many parental clones exhibiting high-level expression of the introduced gene was similar to that of nontransfected dhfr- CHO cells. On the other hand, most parental clones with low-level expression experienced noticeable morphological changes such as bipolar fibroblast-like morphology. Incase of selection of parental clones with TPO expression level higher than200 ng/mL, morphological selection improved selection efficiency by 3.5-fold compared with random selection. Furthermore, when subjected to methotrexate for gene amplification, parental clones that were selected based on morphology elevated the expression level as much as those that were selected randomly. Taken together morphological selection of parental clones can facilitate the establishment of rCHO cell lines expressing recombinant proteins.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 19/01/20 alle ore 00:57:22