Separation by cation-exchange high-performance liquid chromatography of three forms of Chinese hamster ovary cell-derived recombinant human interleukin-2.
Purified recombinant (r) interleukin 2 (IL-2) produced by a transformed Chinese hamster ovary cell line shows a single peak when analysed by reversed-phase high-performance liquid chromatography, but it can be resolved into three forms by sodium dodecyl sulphate polyacrylamide gel electrophoresis. These three forms were successfully isolated by narrow-bore ion-exchange chromatography through optimization of the elution conditions. The addition of n-propanol as an organic modifier to the mobile phase proved to be essential for the recovery of the protein from the column in a yield of 90% or better based on protein quantification and biological activity determination. This chromatographic method was used for the purification of these three rIL-2 forms which represent variable glycosylation of a single polypeptide chain. A comparison of the biological activities using the murine CTLL-2 cell proliferation assay showed that the specific activities of the three forms are similar.
Marchese
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E
Unité Biochimie des Proteines, Sanofi Elf Bio-Recherches, Labège, France.
Vita
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N
Maureaud
T
T
Ferrara
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P
D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D007376:Interleukin-2 / Q000302:isolation & purification*; D051379:Mice; D011994:Recombinant Proteins / Q000302:isolation & purification
D007376:Interleukin-2; D011994:Recombinant Proteins