An ELISA for the measurement of human leukemia inhibitory factor in biological fluids and culture supernatants.

Autor: De Groote D; Department of Endocrinology, University of Liège, Sart Tilman, Belgium., Fauchet F, Jadoul M, Dehart I, Raher S, Gevaert Y, Lopez M, Gathy R, Franssen JD, Radoux D, et. al.
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 1994 Jan 03; Vol. 167 (1-2), pp. 253-61.
DOI: 10.1016/0022-1759(94)90094-9
Abstrakt: A new monoclonal antibody-based ELISA for leukaemia inhibitory factor/human interleukin for DA cells (LIF/HILDA) measurements is described. The sensitivity (56 pg/ml after 4 h incubation, 14 pg/ml after 24 h incubation), precision (intra-assays < 5%), reproducibility (interassay < 10%), and accuracy (recoveries, ranging between 98 and 119%, in several fluids) of the assay, plus its excellent performance in dilution tests, and the lack of interference when in the presence of possible cross-reactive substances guarantee accurate cytokine measurement in biological fluids such as serum, plasma, synovial fluid, follicular fluid, urine and culture supernatants. Using the assay, LIF/HILDA was measurable in supernatants after in vitro whole blood stimulation with phytohemagglutinin (PHA), OKT3, and phorbol myristate acetate (PMA) but not with lipopolysaccharide (LPS) or Ca ionophore. LIF/HILDA production was not measurable until after 24 h of culture, when cytokine levels were seen to increase linearly in the supernatant to reach values of up to 40 ng/ml after 96 h of culture. Finally, a good correlation was found (r = 0.96; p < 0.0001; y = 23.1x + 233) between the LIF/HILDA values obtained using the ELISA and DA-1a bioassay.
Databáze: MEDLINE