Recombinant human extracellular superoxide dismutase produced in milk of transgenic rabbits

• Published in: Transgenic research Vol. 6; no. 4; pp. 271 - 278
• Main Authors: STRÖMQVIST, M, HOUDEBINE, L.-M, ANDERSSON, J.-O, EDLUND, A, JOHANSSON, T, VIGLIETTA, C, PUISSANT, C, HANSSON, L
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.07.1997; Springer Nature B.V; Springer Verlag
• Subjects: Animals; Animals, Genetically Modified; Biological and medical sciences; Biotechnology; CHO Cells - metabolism; Chromatography; Chromatography - methods; Cricetinae; Female; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Glycosylation; Heparin - metabolism; Humans; Life Sciences; Methods. Procedures. Technologies; Milk - enzymology; Peptides - metabolism; Proteins; Rabbits - genetics; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; RNA, Messenger - biosynthesis; Rodents; Superoxide Dismutase - genetics; Superoxide Dismutase - isolation & purification; Superoxide Dismutase - metabolism; Transgenic animals; Transgenic animals and transgenic plants; Enzyme; Transgenic animal; Rabbit; Superoxide dismutase; Lagomorpha; Gene expression; Metalloenzyme; Metalloproteins; Vertebrata; Mammalia; Oxidoreductases; Mammary gland; Recombinant protein; Milk; BIOLOGIE MOLECULAIRE
• ISSN: 0962-8819; 1573-9368
• DOI: 10.1023/A:1018406611380

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.