Expression and characterization of the recombinant swine interleukin-6

• Published in: Comparative immunology, microbiology and infectious diseases microbiology and infectious diseases, 2005-03, Vol.28 (2), p.103-120
• Main Authors: Nuntaprasert, A., Mori, Y., Fujita, K., Yoneda, M., Miura, R., Tsukiyama-Kohara, K., Kai, C.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - immunology; Antibody Formation - drug effects; Anticorps monoclonal; Applied microbiology; Bacterial expression; Baculoviridae - genetics; Baculovirus expression; Biological and medical sciences; Biological Assay - veterinary; Blotting, Western - veterinary; Cercopithecus aethiops; Cloning, Molecular; COS Cells; Cytokine recombinée; DNA, Complementary - genetics; Electrophoresis, Polyacrylamide Gel - veterinary; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Expression bactérienne; Expression baculovilaire; Expression mammaire; Fundamental and applied biological sciences. Psychology; Interleukin-6 - biosynthesis; Interleukin-6 - genetics; Interleukin-6 - immunology; Interleukin-6 - isolation & purification; Mammalian expression; Mice; Microbiology; Polyclonal antibody; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Recombinant Proteins - isolation & purification; Recombinant Swine IL6; Reverse Transcriptase Polymerase Chain Reaction - veterinary; Sequence Analysis, DNA; Spodoptera - metabolism; Spodoptera - virology; Swine - genetics; Swine - immunology; Transfection; Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies (general aspects)
• ISSN: 0147-9571; 1878-1667
• DOI: 10.1016/j.cimid.2004.07.004

The swine interleukin-6 (SwIL-6) cDNA was cloned by RT-PCR and each expression system of recombinant SwIL-6 in Escherichia coli, insect cells, and mammalian cells was developed. Recombinant SwIL-6 produced in bacteria was applied for generation of the polyclonal antibodies. The rSwIL-6 was purified from supernatant of insect cells with a Q-sepharose or anti-SwIL-6 monoclonal antibody based immunoaffinity column. The antibodies showed that the molecular weight of rSwIL-6 was approximately 26 kDa in E. coli, 25, 26, 30 kDa in insect cells, and 26 and 30 kDa in mammalian cells. These variations of molecular weight were probably due to the different modifications of glycosylation. All these recombinant proteins retained the antigenicity and biological activity on 7TD1 mouse cells. Un clone de cDNA de l'interleukine-6 du porc (SwIL-6) a été obtenu par RT-PCR, et des systèmes exprimant la SwIL-6 recombinée ont été exploités dans des cellules d' Escherichia coli d'insectes et de mammifères. La SwIL 6 recombinée provenant des cellules bactérinnes a servi pour préparer l'anticorps polyclonal. La SwIL-6 a été purifiée à partir du surnageant des cellules d'insectes avec la sépharose-Q ou la colonne d'immuno-affinité à l'anticorps monoclonal anti SwIL6. Le poids moléculaire de la SwSIL-6 était de 26 kDa pour les cellules bactériennes, de 25, 26 et 30 kDa pour les cellules d'insectes, et 25 ou 30 kDa pour les cellules de mammifères.