title: Generation and characterization of antibody against porcine interleukin-18.
abstract: The gene-encoding mature porcine interleukin-18 (pIL-18) was amplified by PCR and cloned into a prokaryotic expression vector pET-30a(+). The resulting recombinant plasmid pET-30a-pIL-18 was transformed into Escherichia coli. Expression of recombinant pIL-18 protein was induced by 1 mM isopropyl β-D-thiogalactoside at 37°C. The purified recombinant protein was used to generate a hyperimmune antiserum in a rabbit. The anti-pIL-18 serum was evaluated for its specificity and titer through indirect enzyme-linked immunosorbent assay. The specific reactivity of the anti-pIL-18 antibody was further confirmed by Western blot and immunofluorescence assays. Moreover, the pIL-18 was able to stimulate the proliferation of pig peripheral blood mononuclear cells in vitro, indicating it may have certain biological activity.
authors.affiliation: Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Xiangfang District, Harbin, China.
authors.forename: Siqingaowa
authors.initials: S
authors.lastname: Suo
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Ren
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification / Q000494:pharmacology; D000918:Antibody Specificity; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D005455:Fluorescent Antibody Technique; D020382:Interleukin-18 / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D007544:Isopropyl Thiogalactoside / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D034421:Sus scrofa / Q000235:genetics / Q000276:immunology*
chemical_list: D000906:Antibodies; D020382:Interleukin-18; D011994:Recombinant Proteins; D007544:Isopropyl Thiogalactoside