KMID : 0545119920020020135
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Journal of Microbiology and Biotechnology 1992 Volume.2 No. 2 p.135 ~ p.140
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Partitioning of Recombinant Human Interleukin-2 in a Poly(ethylene glycol)-Dextran Aqueous Two-Phase System
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LEE IN-YOUNG
Lee Sun-Bok
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Abstract
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The partitioning of recombinant human interleukin-2(rhIL-2) in PEG 8000-dextran 38800 aqueous two-phase system has been investigated using three different sources of rhIL-2. In the case of pure rhIL-2, the solubility in a PEG-dextran two-phase system was low and most of rhIL-2 was partitioned into the bottom phase. For the recovery of rhIL-2 from insoluble protein aggregates, the inclusion bodies of recombinant E. coli were solubilized by the treatment with sodium dodecyl sulfate (SDS). The addition of SDS significantly enhanced not only the solubility of rhIL-2 but also the partitioning of rhIL-2 to the top phase. When the ratio of SDS to rhIL-2 was 2.0, the partition coeffident(K) and the recovery yield(Y) at the top phase were 4.5 and 88%, respectively, at pH 6.8. In order to reduce the recovery steps further, SDS was directly added to the intact recombinant E. coli cells and then partitioned into the PEG/dextran aqueous twophase system. The observed partition coefficient (K??3.0) and recovery yield (Y¡Ã80%) of this method were comparable to the rhIL-2 recovery from insoluble protein aggregates. The results obtained in this work indicate that PEG-dextran two-phase partitioning might provide a simple way for the recovery and partial purification of recombinant proteins which are produced as inclusion bodies.
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