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• W1568072008 abstract "Murine interleukin-6 (IL-6), when expressed in Escherichia coli using the pUC9 vector, accumulated as insoluble aggregates or 'inclusion bodies'. After selective urea washing of the inclusion bodies, to remove extraneous proteins, murine IL-6 was solubilized with 8 M guanidine hydrochloride and then rapidly purified to homogeneity by gel-permeation chromatography followed by reversed-phase HPLC. It was demonstrated that complete disulfide bond formation in murine IL-6 occurred during the early urea washing/guanidine hydrochloride extraction steps, so no refolding step was required. When fully reduced murine IL-6 was dissolved in 8 M guanidine hydrochloride and allowed to air-oxidize, complete disulfide bond formation, monitored by analytical reversed-phase HPLC, was shown to occur within 13 h at 6 degrees C. About 25 mg pure protein was obtained from 37 g wet cells. This recombinant murine IL-6 had a specific activity in the hybridoma growth factor assay of 2 x 10(8) U/mg, which is equivalent to that of native murine IL-6. During the purification procedure, a number of variant forms of murine IL-6 were isolated and partially characterized. Two of these forms, T1 and T3, were C-terminal deletants of murine IL-6 lacking about 60 and 20 amino acids from the C-terminus, respectively, while the other form, T2, was an N-terminal deletant lacking 37 amino acids from the N-terminus. None of these variant forms of murine IL-6 bound to the murine IL-6 receptor and, consequently, all were inactive in the hybridoma growth factor assay." @default.
• W1568072008 created "2016-06-24" @default.
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• W1568072008 creator A5036737482 @default.
• W1568072008 creator A5057698507 @default.
• W1568072008 creator A5060396307 @default.
• W1568072008 creator A5090818171 @default.
• W1568072008 date "1992-08-01" @default.
• W1568072008 modified "2023-10-09" @default.
• W1568072008 title "Purification and characterization of a recombinant murine interleukin-6. Isolation of N- and C-terminally truncated forms" @default.
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• W1568072008 doi "https://doi.org/10.1111/j.1432-1033.1992.tb17123.x" @default.
• W1568072008 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/1499565" @default.
• W1568072008 hasPublicationYear "1992" @default.
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