Recombinant human extracellular superoxide dismutase produced in milk of transgenic rabbits |
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Authors: | Stromqvist Mats Houdebine Louis-Marie Andersson Jan-Olof Edlund Anders Johansson Thore Viglietta Celine Puissant Claudine Hansson Lennart |
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Affiliation: | (1) Astra Hassle AB, Tvistev. 48, S-907 36 Umea, Sweden;(2) INRA Unite de Differenciation Cellulaire, 78352 Jouy- en-Josas Cedex, France |
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Abstract: | Expression of human extracellular superoxide dismutase (EC-SOD), a glycosylated, tetrameric metalloprotein, was targeted to the lactating mammary gland of transgenic rabbits. Efficient expression of the recombinant whey acidic protein/ec-sod gene was achieved and up to 3 mg ml–1 of the enzyme was secreted into the milk. Rabbit milk-produced recombinant EC-SOD was primarily found in the whey and purified by a two-step chromatographic method. To evaluate the rabbit milk-produced human EC-SOD, comparisons with native and Chinese hamster ovary cell (CHO)-produced EC-SOD were performed. All proteins were tetrameric and N-glycosylated. The behaviour on SDS-PAGE and size-exclusion chromatography indicated that the masses, and thereby the extent of post-translational modification of the proteins was similar. The monosaccharide composition of both recombinant EC-SOD variants was analysed and indicated similarities in the attached N-glycans on the two proteins. Furthermore, the peptide maps of the three EC-SOD variants revealed that all proteins had similar polypeptide backbones |
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Keywords: | superoxide dismutase recombinant protein transgeneexpression metalloprotein gene expression transgenic rabbit |
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