RESUMO
Highly specific and sensitive immunoassay method for soluble human recombinant interleukin-6 (hu rlL-6) was established by two different immunization methods. One is conventional method by Freund's adjuvant method and the other is special method which is directly injected to mouse spleen. Among seven established monoclonal antibodies (mAbs), two typical monoclonal antibodies, designated YB3 (IgG1) and NY2 (IgM), were further characterized. These mAbs highly bound to IL-6, however did not show cross reactivity with IL-1B and IL-2. As the results of ELISA inhibition assay and western blotting method, it was further identified that YB3 and NY2 had high binding specificity with IL-6. And the limiting detection amount of rlL-6 for YB3 was 5 ng/ml and for NY2 was 0.5 ng/ml. Furthermore, N-glycosylated human rlL-6 was also bound to YB3 on ELISA. On the other hand YB-3 furtherly recognized N-glycosylated human rlL-6 by sandwich ELISA method. These mAbs may be of use to diagnose the gynecopathy which contains abortion and preterm labor.