Turkish Journal of Medical Sciences
Article Title
Development of ELISA Systems for Measurement of Human Tumor Necrosis Factor-Alpha (TNF-\alpha)
DOI
-
Abstract
The aim of this study was to develop a human tumor necrosis factor alpha (hTNF-\alpha) ELISA system because specific and sensitive measurement of low levels of circulating TNF-a is very important for enlightening the immunopathologial mechanisms associated with TNF-\alpha. Monoclonal antibodies 6A4c and 8A6 were produced against hTNF-\alpha and were used as the capture antibody and the tracer antibody respectively for the first hTNF-\alpha ELISA system (6A4c/biotin-8A6). Murine polyclonal IgG was used as the tracer antibody in the second hTNF-\alpha ELISA system (6A4c/biotin-polyclonal IgG). Both systems could detect both recombinant hTNF-\alpha and native hTNF-\alpha. The detection limits defined as minimal concentration of hTNF-\alpha were less than 4 pg/ml for the first and less than 12 pg/ml for the second ELISA systems. 6A4c/biotin-8A6 system resulted in some non-specific reactions to some extent with human sera; however, 6A4c/biotin-polyclonal IgG system produced acceptable background levels with human sera. A prominent inhibitory effect of TNF receptors-I and -II did not occur in any of the ELISA systems at physiological concentrations. Two different types of ELISA systems with high sensitivity and specificity were developed to measure hTNF-\alpha level both in human serum and cell culture supernatant.
First Page
385
Last Page
393
Recommended Citation
YÜCEL, AYŞEGÜL and AYBAY, CEMALETTİN (2005) "Development of ELISA Systems for Measurement of Human Tumor Necrosis Factor-Alpha (TNF-\alpha)," Turkish Journal of Medical Sciences: Vol. 35: No. 6, Article 6. Available at: https://journals.tubitak.gov.tr/medical/vol35/iss6/6