an evaluation of commercial fluorescent bead-based luminex cytokine assays评估商业荧光bead-based luminex细胞因子分析.pdfVIP

An Evaluation of Commercial Fluorescent Bead-Based Luminex Cytokine Assays 1 1 1 1 1 Joel Fleury Djoba Siawaya *, Teri Roberts , Chantal Babb , Gillian Black , Hawa Jande Golakai , Kim 1 1 1 2 1 Stanley , Nchinya Bennedict Bapela , Eileen Hoal , Shreemanta Parida , Paul van Helden , Gerhard Walzl1 1 Department of Biomedical Sciences, Division of Molecular Biology and Human Genetics, Stellenbosch University Cape-Town, Stellenbosch, South Africa, 2 Department of Immunology, Max Planck Institute for Infection Biology, Berlin, Germany Abstract The recent introduction of fluorescent bead-based technology, allowing the measurement of multiples analytes in a single 25–50 ml sample has revolutionized the study of cytokine responses. However, such multiplex approaches may compromise the ability of these assays to accurately measure actual cytokine levels. This study evaluates the performance of three commercially available multiplex cytokine fluorescent bead-based immunoassays (Bio-Rad’s Cytokine 17-plex kit; LINCO Inc’s 29-plex kit; and RnD System’s Fluorokine-Multi Analyte Profiling (MAP) base kit A and B). The LINCO Inc kit was found to be the most sensitive assay for measuring concentrations of multiple recombinant cytokines in samples that had been spiked with serial dilutions of the standard provided by the manufacturer, followed respectively by the RnD Fluorokine- (MAP) and Bio-Rad 17-plex kits. A positive correlation was found in the levels of IFN-c measured in antigen stimulated whole blood culture supernatants by the LINCO Inc 29-plex, RnD Fluorokine-(MAP) and RnD system IFN-c Q