[Development of sandwich ELISA for equine interferon-gamma detection].
OBJECTIVE
To develop a quantitative ELISA by measuring interferon (IFN-gamma) of equine lymphocytes.


METHODS
Sandwich ELISA for equine IFN-gamma was developed using mAb A5 as a capture antibody and biotinylated mAb SB10 as a detection antibody.


RESULTS
The detection limit of the sandwich ELISA for equine IFN-gamma was 1 microg/L and did not show cross-reactivity with recombinant equine IL-18. Equine IFN-gamma was detected by ELISA in culture medium of the peripheral blood mononuclear cells (PBMCs) stimulated with ConA or PMA/Ionomycin.


CONCLUSIONS
This method can be used to help understand the role of this cytokine in various equine diseases and develop specific cell-mediated immunity assay.
Bai
Yu
Y
National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute CAAS, Harbin 150001, China. baiyu106@163.com
Tong
Tie-gang
TG
Zhang
Wei-jun
WJ
Xu
Shu-lan
SL
Wang
Qun
Q
Liu
Guang-liang
GL
Wu
Dong-lai
DL
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001681:Biological Assay; D016207:Cytokines / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006736:Horses; D007371:Interferon-gamma / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D020382:Interleukin-18 / Q000276:immunology / Q000302:isolation & purification*; D015759:Ionomycin / Q000737:chemistry; D057230:Limit of Detection; D008213:Lymphocyte Activation; D009500:Neutralization Tests; D011994:Recombinant Proteins; D012685:Sepharose / Q000031:analogs & derivatives / Q000737:chemistry; D050378:T-Lymphocytes, Regulatory
D000911:Antibodies, Monoclonal; D016207:Cytokines; D020382:Interleukin-18; D011994:Recombinant Proteins; C028326:concanavalin A-sepharose; D015759:Ionomycin; D007371:Interferon-gamma; D012685:Sepharose