Characterization of recombinant glycosylated human interleukin 2 produced by a recombinant plasmid transformed CHO cell line
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...
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Veröffentlicht in: | FEBS letters 1987-12, Vol.226 (1), p.47-52 |
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Sprache: | eng |
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Zusammenfassung: | 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 × 10
7 U/mg. |
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ISSN: | 0014-5793 1873-3468 |
DOI: | 10.1016/0014-5793(87)80548-3 |