title: High-level expression of a biologically active human interleukin-6 mutein.
abstract: We have constructed two different muteins of interleukin-6 (IL-6) which were expressed in Escherichia coli. Both muteins lack the first 22 N-terminal amino acids of native IL-6 and lack one or the other of the two naturally occurring pairs of cysteines at either position 45 and 51 or position 74 and 84 of IL-6. We found that there was a dramatic increase in the level of IL-6 produced from each mutein clone, compared to the level produced by the wild-type IL-6 clone. We also observed that the yield of soluble and properly refolded mutein IL-6 was highest when the cysteines at position 74 and 84 were left intact. The mutein IL-6 with cysteines at position 74 and 84 was as active as wild-type IL-6 and a lower concentration of the mutein IL-6 was required to reach maximal activity, compared to wild-type IL-6. The mutein IL-6 with cysteines at position 45 and 51 had a much reduced biological activity.
authors.affiliation: ImClone Systems Inc., New York, NY 10014.
authors.forename: S M
authors.initials: SM
authors.lastname: Skelly
authors.forename: C
authors.initials: C
authors.lastname: Tackney
authors.forename: D
authors.initials: D
authors.lastname: Hicklin
authors.forename: T
authors.initials: T
authors.lastname: Tamkins
authors.forename: N
authors.initials: N
authors.lastname: Goldstein
authors.forename: H
authors.initials: H
authors.lastname: Waksal
authors.forename: S
authors.initials: S
authors.lastname: Dagan
mesh_terms: D000818:Animals; D006801:Humans; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D051379:Mice; D009154:Mutation; D013329:Structure-Activity Relationship
chemical_list: D015850:Interleukin-6