A method for correcting for the variability of inhibitory effects of soluble human interleukin 1 receptor II measured by different ELISAS.
Seven ELISAs were developed by using several combinations of anti-human IL-1beta antibodies for detecting interleukin 1beta (IL-1beta) in cell culture supernatants. These ELISAs have different sensitivities in detecting standard preparations of recombinant human IL-1beta (WHO reference standard) compared with conventional preparations of IL-1beta produced by stimulated human peripheral blood mononuclear cells. The observed differences were attributed to differences in epitope specificity of the various monoclonal antibodies used and the heterogeneity of IL-1beta secreted into culture supernatants. The presence of soluble IL-1 receptor type I did not alter the levels of IL-1beta detected by these ELISAs. However, soluble IL-1 receptor type II interfered with the detection of IL-1beta to different degrees in these ELISAs. A method involving standarization by means of separate measurement of the amount of receptor and its inhibitory effect in the IL-1beta ELISA, yields consistent estimates of the correct IL-1beta levels.
Krakauer
T
T
Department of Immunology and Molecular Biology, United States Army Medical Research Institute of Infectious Diseases, Frederick, MD 21702-5011, USA.
Krakauer
H
H
D016477:Artifacts; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007375:Interleukin-1 / Q000032:analysis*; D017472:Receptors, Interleukin-1; D053574:Receptors, Interleukin-1 Type II; D012015:Reference Standards; D015203:Reproducibility of Results; D012995:Solubility
D007375:Interleukin-1; D017472:Receptors, Interleukin-1; D053574:Receptors, Interleukin-1 Type II