title: Establishment of a quantitative bovine CXCL8 sandwich ELISA with newly developed monoclonal antibodies.
abstract: Three IgG class anti-bovine CXCL8 (bCXCL8) monoclonal antibody (mAb)-secreting hybridomas, SH8-8D7, SH8-12A5 and SH8-2A1, were developed. SH8-8D7 was IgG2a, and SH8-12A5 and SH8-2A1 were IgG1. All three mAbs detected recombinant bCXCL8 (rbCXCL8) by immunoprecipitation and Western blotting. SH8-2A1 could neutralise the chemotactic activity of rbCXCL8 towards neutrophils. The quantitative bCXCL8 ELISA was constituted by the combination of SH8-12A5 and biotin-SH8-2A1. The detection range was 20-1000  pg/mL. A sandwich ELISA was used to measure native bCXCL8 derived from the supernatant of cultured bovine peripheral blood mononuclear cells stimulated with ConA, LPS or PHA. Furthermore, SH8-2A1 could detect bCXCL8 in formalin-fixed, paraffin-embedded, pneumonic calf tissues. These findings indicate that the newly developed anti-bCXCL8 mAbs could contribute to research on bovine inflammatory responses and immunology.
authors: The Research Team for Advanced Biologicals, National Institute of Animal Health, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan.
authors: Jiro
authors: J
authors: Hirota
authors: Shinya
authors: S
authors: Shimizu
authors: Atsushi
authors: A
authors: Watanabe
authors: Fumiko
authors: F
authors: Suzuta
authors: Kazue
authors: K
authors: Yajima
authors: Kumiko
authors: K
authors: Kimura
authors: Makoto
authors: M
authors: Haritani
authors: Shigeki
authors: S
authors: Inumaru
authors: Yukio
authors: Y
authors: Yagi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002417:Cattle; D003429:Cross Reactions / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007150:Immunohistochemistry; D016209:Interleukin-8 / Q000276:immunology*; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000276:immunology; D012015:Reference Standards; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D016209:Interleukin-8; D011994:Recombinant Proteins