Chicken IFN-gamma monoclonal antibodies and their application in enzyme-linked immunosorbent assay.
Twelve mabs against native or recombinant chicken IFN-gamma were produced and characterized by virus neutralization, ELISA, and Western blot assays. No data were obtained to suggest that the form of the immunogen (native versus recombinant) influenced the antigenic specificity of the mabs produced. While only two antibodies inhibited the in vitro virus neutralizing activity of IFN-gamma, other evidence indicated that the specificity of these mabs was indeed directed against IFN-gamma. By Western blot analysis, all antibodies identified a 17-kDa IFN-gamma polypeptide. Using a direct binding ELISA incorporating these mabs, a high correlation with IFN-gamma detected by in vitro virus neutralization was observed. The IFN-gamma ELISA was also capable of measuring cytokine levels in the sera of chickens orally infected with Eimeria maxima. At 8 and 10 days post-primary infection, significantly higher (p<0. 001) levels of serum IFN-gamma were detected in E. maxima infected chickens compared to uninfected controls. These results indicate that a mab-based direct binding ELISA is suitable to measure chicken IFN-gamma in a variety of formats.
Yun
C H
CH
BARC-East, Building 1040, Immunology and Disease Resistance Laboratory, Livestock and Poultry Sciences Institute, USDA, Agricultural Research Service, Beltsville, MD 20705, USA.
Lillehoj
H S
HS
Choi
K D
KD
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000096:biosynthesis; D000918:Antibody Specificity; D015153:Blotting, Western / Q000662:veterinary; D002645:Chickens / Q000276:immunology*; D003048:Coccidiosis / Q000276:immunology / Q000469:parasitology / Q000662:veterinary*; D016207:Cytokines; D004539:Eimeria; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D007371:Interferon-gamma / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008934:Mitogens / Q000494:pharmacology; D009500:Neutralization Tests / Q000662:veterinary; D011201:Poultry Diseases / Q000276:immunology* / Q000469:parasitology; D011994:Recombinant Proteins; D013154:Spleen / Q000187:drug effects
D000911:Antibodies, Monoclonal; D016207:Cytokines; D008934:Mitogens; D011994:Recombinant Proteins; D007371:Interferon-gamma