Characterization of monoclonal antibodies against goat interleukin-18 and their application in the measurement of goat interleukin-18 in LPS-stimulated peripheral blood mononuclear cells by sandwich ELISA.
In order to develop a specific assay for the measurement of goat IL-18 level, two stable hybridoma cell lines were established which secreted IgG1 monoclonal antibodies (mAbs) against goat IL-18. Specific binding of two mAbs named 2E8 and 4C4 to recombinant goat IL-18 expressed in Escherichia coli was demonstrated in an ELISA and Western blotting. Results also showed that mAbs 2E8 and 4C4 bound to distinct epitopes in the ELISA additivity test. These two mAbs were applied in IFA analysis for the detection of goat IL-18 expressed in 293FT cells and in the sandwich ELISA for the measurement of goat IL-18 levels in LPS-stimulated PBMC. Results from this study demonstrated that mAbs against goat IL-18 recognize bovine and human IL-18 and could be used to measure IL-18 levels in different inflammations or immune responses in future studies.
Cao
Bing-lei
BL
Shandong Agricultural University, Taian 271018, China.
Qiu
Hong-kai
HK
Hu
Jing-dong
JD
Meng
Fan-dong
FD
Zhang
Hong
H
Zhao
Hong-kun
HK
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods / Q000662:veterinary; D006041:Goats / Q000276:immunology*; D057809:HEK293 Cells; D006801:Humans; D066298:In Vitro Techniques; D020382:Interleukin-18 / Q000032:analysis* / Q000235:genetics / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D020382:Interleukin-18; D008070:Lipopolysaccharides; D011994:Recombinant Proteins