Production of Polyclonal Antibody against Interleukin-33 and Assessment of Its Distribution in Murine Liver and Lung

• Published in: BioMed research international 2009-01, Vol.2009 (2009), p.1-6
• Main Authors: Liu, Xiaojin, Wu, Yan, Li, Mingcai, Chen, Sui, Zhou, Yanchun
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - immunology; Asthma; Bacteria; Biological and medical sciences; Cells, Cultured; E coli; Escherichia coli; Gene expression; Identification and classification; Interleukin-33; Interleukins; Interleukins - immunology; Kinases; Liver; Liver - immunology; Lung - immunology; Medical sciences; Methods; Mice; Mice, Inbred BALB C; Pharmacology. Drug treatments; Physiological aspects; Polyclonal antibodies; Protein Engineering - methods; Rabbits; Recombinant Proteins - immunology; Recombinant Proteins - metabolism; Rheumatoid arthritis; Signal transduction; Tissue Distribution
• ISSN: 1110-7243; 2314-6133; 1110-7251; 2314-6141
• DOI: 10.1155/2009/729197

Interleukin (IL)-33 is the latest member of IL-1 cytokine family. In this study, the cloning, expression, purification, and polyclonal antibody preparation of mouse IL-33 were described. The coding region of IL-33 mature protein was cloned into the prokaryotic expression vector pET-44. The recombinant protein, IL-33 containing a hexahistidine tag in the C-terminal, was expressed in Escherichia coli. The expressed soluble protein was purified by immobilized metal-ion affinity chromatography using Ni2+-nitrilotriacetic acid agarose. The rabbits were immunized with the purified recombinant protein. The obtained antiserum was precipitated by saturated ammonium sulfate and then purified by Protein A affinity chromatography. The sensitivity and specificity of the antibodies were evaluated by enzyme-linked immunosorbent assay and immunohistochemistry. The high titer (1:32000) polyclonal antibodies with high specificity were obtained by immunizing rabbits with the purified recombinant protein. Significant expression of IL-33 was seen in mouse liver and lung tissues determined with the anti-IL-33. The production of the polyclonal antibody against IL-33 provides a good tool for studying the biofunctions of IL-33.