The production and characterization of anti-bovine CD14 monoclonal antibodies

Veterinary Research
Eun J SohnDouglas D Bannerman

Abstract

To characterize further the chemical and biological properties of bovine soluble (bos) CD14, a panel of ten murine monoclonal antibodies (mAb) reactive with recombinant (r) bosCD14 were produced. A sandwich ELISA, using murine mAb and rabbit polyclonal antibodies reactive with rbosCD14 was developed. All the mAb were reactive by ELISA with baculovirus-derived rbosCD14 and they recognized rbosCD14 (40 kDa) by western blot analysis. The mAb also identified by western blot sCD14 (53 and 58 kDa) in milk and blood and sCD14 (47 kDa) in a lysate of macrophages obtained from involuted bovine mammary gland secretions. Analysis by ELISA of whey samples after intramammary injection of lipopolysaccharide (LPS) (10 micro g) revealed increased sCD14 levels between 8 to 48 h after injection. Flow cytometric analysis showed that the mAb bound to macrophages isolated from involuted mammary gland secretions and mouse macrophages but not to swine or horse monocytes. Addition of anti-rbosCD14 mAb to monocytes stimulated with LPS reduced in vitro production of TNF-alpha. The anti-rbosCD14 antibodies generated in this study will be useful in studying CD14, an accessory molecule that contributes to host innate recognition of bacterial cell wall comp...Continue Reading

Citations

Sep 4, 2007·American Journal of Veterinary Research·Cleverson D SouzaDouglas J Weiss
Jan 16, 2010·Veterinary Research Communications·Yoshio KikuTomohito Hayashi
Apr 14, 2006·Veterinary Research·Pascal Rainard, Céline Riollet
Jul 4, 2006·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Lev G NemchinovRosemarie W Hammond
Oct 16, 2013·British Journal of Haematology·Cécile Durousseau de CoulgeansSylvie Chapel-Fernandes

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