[Immunoenzyme determination of human granulocyte-macrophage colony-stimulating factor using monoclonal antibodies].
A set of seven hybridomas producing monoclonal antibodies (MAbs) to the human recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) was obtained. The properties of the monoclonal antibodies were characterized, and pairs of MAbs specific to different non-overlapping epitopes of GM-CSF were identified. A sensitive and simple method of two-site ELISA for GM-CSF was developed on the basis of two MAbs. According to this method, one MAb is absorbed onto a microtiter plate and another is labeled with biotin and used for the detection of GM-CSF bound to the first MAb. MAb labeled with biotin, in its turn, was visualized with the streptavidin-horseradish peroxidase conjugate. The sensitivity of this test was no less than 0.5 ng/ml, and a linear dose-response relationship was observed within a concentration interval from 0.5 to 32 ng/ml. No cross-reactivity was found with human tumor necrosis factor-alpha, granulocyte colony-stimulating factor, interleukin-2, or interleukin-3 in this test system.
Lakhtina
O E
OE
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
Petrovskaia
L E
LE
Korobko
V G
VG
Nemeianov
V A
VA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D003429:Cross Reactions; D016179:Granulocyte Colony-Stimulating Factor / Q000276:immunology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis* / Q000276:immunology; D006801:Humans; D007376:Interleukin-2 / Q000276:immunology; D007377:Interleukin-3 / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D012680:Sensitivity and Specificity; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
D000911:Antibodies, Monoclonal; D007376:Interleukin-2; D007377:Interleukin-3; D014409:Tumor Necrosis Factor-alpha; D016179:Granulocyte Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor