Variability in the sensitivity of nine enzyme-linked immunosorbant assays (ELISAs) in the measurement of human interleukin 6.
ELISAs employing several combinations of polyclonal and monoclonal anti-human IL-6 antibodies for the detection of interleukin 6 (IL-6) in cell culture supernatants were developed and compared. They were found to have differing sensitivities in the measurement of standard preparations of recombinant human (rh) IL-6 (WHO reference standard) as well as conventional preparations of IL-6 produced by stimulated human peripheral blood mononuclear cells. Thus, an ELISA that was optimal for detecting rhIL-6 standard was suboptimal for detecting IL-6 in cell culture supernatants. The presence of soluble IL-6 receptor (sIL-6R) or the signal-transducing gp130 molecule or serum did not alter the levels of IL-6 detected by these ELISAs. Therefore, the observed variability in sensitivity of the assays may be due to differences in epitope specificity of the various monoclonal antibodies used and the heterogeneity of IL-6 secreted into culture supernatants.
Krakauer
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Department of Immunology and Molecular Biology, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702-5011, USA.
D000918:Antibody Specificity; D015703:Antigens, CD / Q000032:analysis; D001798:Blood Proteins / Q000032:analysis; D002478:Cells, Cultured; D050822:Cytokine Receptor gp130; D004768:Enterotoxins / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D015850:Interleukin-6 / Q000032:analysis* / Q000276:immunology; D007963:Leukocytes, Mononuclear; D008562:Membrane Glycoproteins / Q000032:analysis; D019947:Receptors, Interleukin-6 / Q000032:analysis; D011994:Recombinant Proteins / Q000276:immunology; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012995:Solubility; D018089:Superantigens / Q000276:immunology; D014944:World Health Organization
D015703:Antigens, CD; D001798:Blood Proteins; D004768:Enterotoxins; C494199:IL6ST protein, human; D015850:Interleukin-6; D008562:Membrane Glycoproteins; D019947:Receptors, Interleukin-6; D011994:Recombinant Proteins; D018089:Superantigens; D050822:Cytokine Receptor gp130; C031818:enterotoxin B, staphylococcal