Characterization of mouse monoclonal antibodies to human interferon-gamma.
Mouse monoclonal antibodies (mAb) to human interferon-gamma (HuIFN-gamma) were characterized. The mAbs studied--E4-18, G4-15, and SAT-1--which are all IgGl-type, reacted to all HuIFN-gamma molecular species, both glycosylated and non-glycosylated. Affinity constants calculated of E4-18 and G4-15 didn't have considerable differences for both kinds of HuIFN-gamma (1-3 x 10(8) liter/mol), but SAT-1 had a difference--a higher value (10(10) liter/mol) for the former than for the latter (8 x 10(8) liter/mol). In epitope specificity, the results suggested that E4-18 and G4-15 recognized an overlapped region remote from the region of SAT-1. Competition experiment using synthetic peptides suggested that epitope of G4-15 is around N9-26 of the HuIFN-gamma sequence. Those mAbs could be used for sandwich radioimmunoassay of HuIFN-gamma using double mAbs in two combinations, one (G4-15/E4-18) based on dimer forms of HuIFN-gamma and the other (SAT-1/E4-18) based on epitope difference. The mAbs are all neutralizing antibodies in which SAT-1 neutralized at a lower concentration than did G4-15, and at a much lower one than did E4-18. The receptor binding of HuIFN-gamma was inhibited by mAbs G4-15 and SAT-1. Efficacy of G4-15 and SAT-1 for the inhibition correspond with that for neutralization.
Yamamoto
Y
Y
Department of Molecular and Cellular Virology, Kyoto University.
Miyata
K
K
Ueda
M
M
Kawade
Y
Y
Matsumoto
K
K
Tsukui
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D011863:Radioimmunoassay; D011971:Receptors, Immunologic / Q000378:metabolism; D017471:Receptors, Interferon
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011971:Receptors, Immunologic; D017471:Receptors, Interferon; D007371:Interferon-gamma