A sandwich enzyme-linked immunosorbent assay for human interleukin-5.
A sandwich enzyme-linked immunosorbent assay (ELISA) was developed for human interleukin-5 (hIL-5) using a combination of monoclonal anti-recombinant(r)-hIL-5 antibody and rabbit anti-r-hIL-5 IgG. Detection limit of this assay was estimated to be 7.8 pg/ml, which was about 10,000 times more sensitive than that of the bioassay using BCL1 cells of murine origin. This ELiSA was specific for hIL-5, showing no crossreactivity to recombinant human GM-CSF, IL-4, TNF-alpha, IFN-gamma and mouse IL-5 (mIL-5). The presence of 10% fetal calf serum did not interfere with the measurement of r-hIL-5. Coefficients of variation in intra-assay and interassay were 1.1-4.6% and 2.3-11.3%, respectively. These results indicate that this assay system can be quite useful in quantifying hIL-5 in various biological fluids.
Suntory Institute for Biomedical Research, Osaka, Japan.
Y
Y
Fukuda
J
J
Hashino
M
M
Haruyama
N
N
Tsuruoka
H
H
Nakazato
T
T
Nakanishi
D000818:Animals; D000911:Antibodies, Monoclonal; D006224:Cricetinae; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D015848:Interleukin-5 / Q000032:analysis*; D007945:Leukemia, Lymphoid / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis; D012680:Sensitivity and Specificity; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha
D000911:Antibodies, Monoclonal; D015848:Interleukin-5; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha