title: A time-resolved fluorescence immunoassay (DELFIA) increases the sensitivity of antigen-driven cytokine detection.
abstract: In an effort to improve the quantification of the low levels of cytokines released in response to antigenic stimulation of T cells, a sandwich dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA) was developed and compared to a standard sandwich ELISA. The DELFIA enhanced the sensitivity of a mouse IL-2 assay 8- to 27-fold, and a human GM-CSF assay 10-fold, as compared to colorimetric ELISA. The increase in sensitivity allows for the use of lower sample volumes per well, and the ability to run more assays per supernatant sample. This sensitive, nonisotopic alternative to other cytokine detection methods will be useful for those researchers wanting to quantitate low levels of antigen-driven cytokine production.
authors.affiliation: Torrey Pines Institute for Molecular Studies, San Diego, California 92121, USA.
authors.forename: Gina
authors.initials: G
authors.lastname: Allicotti
authors.forename: Eva
authors.initials: E
authors.lastname: Borras
authors.forename: Clemencia
authors.initials: C
authors.lastname: Pinilla
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D003470:Culture Media / Q000737:chemistry; D016207:Cytokines / Q000032:analysis / Q000096:biosynthesis* / Q000592:standards; D004797:Enzyme-Linked Immunosorbent Assay; D005063:Europium; D015200:Fluoroimmunoassay / Q000379:methods* / Q000592:standards; D016179:Granulocyte Colony-Stimulating Factor / Q000032:analysis; D006801:Humans; D007376:Interleukin-2 / Q000032:analysis; D051379:Mice; D015203:Reproducibility of Results; D013601:T-Lymphocytes / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis
chemical_list: D000911:Antibodies, Monoclonal; D003470:Culture Media; D016207:Cytokines; D007376:Interleukin-2; D014409:Tumor Necrosis Factor-alpha; D016179:Granulocyte Colony-Stimulating Factor; D005063:Europium