Characterization of recombinant glycosylated human interleukin 2 produced by a recombinant plasmid transformed CHO cell line

FEBS Letters

Ferrara, P.¹  Roskam, W.²  Marchese, E.¹  Vita, N.¹  Lupker, J.²  Pecceu, F.¹ 

[1] Unité Biochimie des Proteines, Sanofi Elf Bio-Récherches, BP 137, 31328 Labege Cedex, France;Unité Technologie des Cellules Animales Recombinées, Sanofi Elf Bio-Récherches, BP 137, 31328 Labege Cedex, France

关键词: Glycoprotein;    Interleukin 2;    Recombinant lymphokine;   

DOI  :  10.1016/0014-5793(87)80548-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A recombinant plasmid containing expression units for human pre-interleukin 2 (pre-IL-2) and the selectable marker mouse DHFR, was constructed and used to transform DHFR⁻ CHO cells to the DHFR⁺ phenotype. Selected colonies were isolated and tested for IL-2 production. Twelve highly IL-2-producing clones were amplified in stepwise increasing concentrations of methotrexate. The IL-2 secreted into the culture medium by one of these clones was purified to homogeneity and partially characterized. N-terminal sequence analysis showed that pre-IL-2 was correctly processed during secretion. SDS gel electrophoresis and chromatofocusing experiments in conjunction with neuraminidase treatment indicated a posttranslational glycosylation of the secreted mature protein similar to that described for the tetrasaccharide structure of the N2 form of natural IL-2. This recombinant IL-2 has a specific activity of 2.5 × 10⁷ U/mg.

【 授权许可】

Unknown