Determination of human tumor necrosis factor alpha by a highly sensitive enzyme immunoassay.
Tumor necrosis factor alpha (TNF-alpha) is a polypeptide produced primarily by monocytes and macrophages. It is involved in a wide variety of immune reactions. A simple and sensitive microplate enzyme-linked immunosorbent assay for the detection of hTNF-alpha in serum, plasma, and cell culture supernatants is described. The method is based on the use of horseradish peroxidase in biotin-streptavidin amplification system which is performed in Nunc StarWell. This system has enabled us to achieve a sensitivity of 0.1 pg hTNF-alpha/ml of the sample. The assay is calibrated to the World Health Organization (WHO) standard for hTNF-alpha (87/650). The within-run coefficient of variation ranged from 3.7 to 5.9 and the between-run coefficient of variation ranged from 8.0 to 9.9. The results obtained by the proposed method and by a commercially available kit (DRG hTNF-alpha ELISA) correlated well (n = 20, r = 0.956).
Hedayati
M
M
Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Islamic Republic of Iran.
Yazdanparast
R
R
Azizi
F
F
D001710:Biotin; D001774:Blood Chemical Analysis / Q000379:methods / Q000592:standards / Q000706:statistics & numerical data; D004797:Enzyme-Linked Immunosorbent Assay; D006735:Horseradish Peroxidase; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D012015:Reference Standards; D012680:Sensitivity and Specificity; D019809:Streptavidin; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis* / Q000592:standards
D014409:Tumor Necrosis Factor-alpha; D001710:Biotin; D019809:Streptavidin; D006735:Horseradish Peroxidase