TY  - EJOU
AU  - Hirota, Jiro 
AU  - Shimizu, Shinya 
AU  - Watanabe, Atsushi 
AU  - Suzuta, Fumiko 
AU  - Yajima, Kazue 
AU  - Kimura, Kumiko 
AU  - Haritani, Makoto 
AU  - Inumaru, Shigeki 
AU  - Yagi, Yukio 

TI  - Establishment of a quantitative bovine CXCL8 sandwich ELISA with newly developed monoclonal antibodies
T2  - European Cytokine Network

PY  - 2011
VL  - 22
IS  - 1
SN  - 1952-4005

AB  - 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 antibCXCL8
mAbs could contribute to research on bovine inflammatory responses and immunology.
KW  - bovine
KW  -  CXCL8
KW  -  monoclonal antibody
KW  -  immunohistochemistry
KW  -  sandwich ELISA

DO  - 10.1684/ecn.2011.0275