Expression and characterization of the recombinant swine interleukin-6.
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 26kDa in E. coli, 25, 26, 30kDa in insect cells, and 26 and 30kDa 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.
Nuntaprasert
A
A
Laboratory of Animal Research Center, Institution of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan.
Mori
Y
Y
Fujita
K
K
Yoneda
M
M
Miura
R
R
Tsukiyama-Kohara
K
K
Kai
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000917:Antibody Formation / Q000187:drug effects; D016367:Baculoviridae / Q000235:genetics; D001681:Biological Assay / Q000662:veterinary; D015153:Blotting, Western / Q000662:veterinary; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D051379:Mice; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D017422:Sequence Analysis, DNA; D018411:Spodoptera / Q000378:metabolism / Q000821:virology; D013552:Swine / Q000235:genetics* / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D015850:Interleukin-6; D011994:Recombinant Proteins