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DELFIA fluorescent immunoassay

Pro-inflammatory cytokines are important mediators of inflammation and tissue destruction. This section describes two cell-based assays that were used to screen for inhibitors of cytokine production and some of the compounds discovered using these screens. The two screens were important elements of a collaboration between Xenova Ltd and the Suntory Institute of Biomedical Research to find microbial metabolites with potential utility for the treatment of rheumatoid arthritis. Both screens were cell stimulatory assays with similar formats, the principle of which is illustrated in Figure 3. Treatment of cells with a particular stimulus activates a signal transduction pathway, one of the end results of which is production of a cytokine, which is secreted into the assay medium. After a separation step, the cytokine of interest is measured quantitatively in the supernatant by dissociation enhanced lanthanide fluorescence immunoassay (DELFIA) using a europium-labeled tertiary antibody. At the same time, cytotoxic properties of test substances are determined by assessing their effect on proliferation of the separated cells. [Pg.90]

II assay in plasma and urine of human volunteers by dis- 48. sociation enhanced lanthanide fluorescence immunoassay (DELFIA). J. Pharm. Biomed. Anal. 1998,16 (5), 883-792. [Pg.1579]

The original inhibition test has been replaced by an immunoradiometric assay and a time-resolved fluorescent immunoassay (the DELFIA method developed by Pharmacia, Uppsala, Sweden). The cut-off values for healthy subjects vary from 14 to 20 kU/L, depending on the method. [Pg.773]

Different variations of time-resolved luminescence assays were patented in 1982 by Wieder [1], in 1983 by Soini and HemmUa [2], and in 1999 by Diamandis [3]. The Finnish company Wallac first commercialized the principle and introduced an assay reader for dissociation-enhanced lanthanide fluorescence immunoassay (DELFIA) in the beginning of the 1980s. The first (1982) DELFIA-based bioassay for diagnostic market was for Rubella antibodies, and it was the first sensitive nonradioisotope innnunoassay marking the beginning of a new era [4]. [Pg.264]

Allicotti, G., Borras, E., and Pinilla, C. (2003) A time resolved fluorescence immuno assay (DELFIA) increases the sensitivity of antigen driven cytokine detection. Journal of Immunoassay and Immunochemistry, 24, 345 358. [Pg.367]

The use of fluorescent labels has been very successful for inorganic complexes such as the europium chelates, which are used today in the Delfia commercial system. In contrast, fluorescence has rarely been employed in the area of organometallic tracers, and only the immunoassay developed by Lakowicz uses such a tracer [88]. This is a homogeneous competitive immunoassay, the tracer being the complex (Re-L) -HSA, 48 obtained as shown in Scheme 8.20 and the detection method fluorescence polarization (FP). This compound 48 displays highly polarized emission (with a maximum polarization near 0.4 and maximum anisotropy near 0.3) in the absence of rotational diffusion and a long average lifetime (2.7 ps) when bound to proteins in air-equilibrated aqueous solution. [Pg.293]

Figure 9.3 Comparison data of fluorescence detection to elemental detection of an hCG immunoassay. Through collaboration with North York General Hospital, the DELFIA system was compared directly with ICP-MS detection. In this assay, DELFIA sandwich ELISA for patient hCG was quantitated as directed by the manufacturer using Eu-tagged anti-hCG antibodies. After analysis using the DELFIA fluorometer, the assay was diluted for elemental analysis by ICP-MS with 50 pL concentrated HCI containing an internal standard of 1 ppb Ho. Cal n stds calibration standards low med hi stds low, medium, and high standards. Figure 9.3 Comparison data of fluorescence detection to elemental detection of an hCG immunoassay. Through collaboration with North York General Hospital, the DELFIA system was compared directly with ICP-MS detection. In this assay, DELFIA sandwich ELISA for patient hCG was quantitated as directed by the manufacturer using Eu-tagged anti-hCG antibodies. After analysis using the DELFIA fluorometer, the assay was diluted for elemental analysis by ICP-MS with 50 pL concentrated HCI containing an internal standard of 1 ppb Ho. Cal n stds calibration standards low med hi stds low, medium, and high standards.

See other pages where DELFIA fluorescent immunoassay is mentioned: [Pg.399]    [Pg.87]    [Pg.331]    [Pg.399]    [Pg.2031]    [Pg.466]    [Pg.967]    [Pg.173]    [Pg.641]    [Pg.344]    [Pg.345]    [Pg.345]    [Pg.335]    [Pg.1237]    [Pg.332]    [Pg.286]    [Pg.747]    [Pg.229]   


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