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.
机构:
USDA ARS, GRAND FORKS HUMAN NUTR RES CTR, POB 9034, UNIV STN, GRAND FORKS, ND 58202 USAUSDA ARS, GRAND FORKS HUMAN NUTR RES CTR, POB 9034, UNIV STN, GRAND FORKS, ND 58202 USA
ROSSOW, KL
REEVES, PG
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机构:
USDA ARS, GRAND FORKS HUMAN NUTR RES CTR, POB 9034, UNIV STN, GRAND FORKS, ND 58202 USAUSDA ARS, GRAND FORKS HUMAN NUTR RES CTR, POB 9034, UNIV STN, GRAND FORKS, ND 58202 USA