title: Multiplexed Microsphere Suspension Array-Based Immunoassays.
abstract: ELISA is an extremely powerful tool to detect analytes because of its sensitivity, selectivity, reproducibility and ease of use. Here we describe sandwich immunoassays performed in suspension on spectrally unique microspheres developed by Luminex. Luminex assays offer the benefit of multiplex analysis of large numbers of analytes in a single reaction. Because the microspheres are spectrally unique, many microspheres, each attached to various antibodies, can be added to a single sample. Luminex instruments can distinguish each microsphere and detect the intensity of a reporter signal for each microsphere. Results are reported in Median Fluorescent Intensities for each analyte. Luminex assays can be used to detect up to 500 analytes in a high-throughput format. Luminex refers to this technology as xMAP(®). Here we describe a routine protocol for a Luminex immunoassay. Other Luminex assays would have to be optimized for specific conditions according to their use.
authors.affiliation: United States Food and Drug Administration (FDA), 1431 Harbor Bay Parkway, Alameda, CA, 94502, USA, andrew.lin@fda.hhs.gov.
authors.forename: Andrew
authors.initials: A
authors.lastname: Lin
authors.forename: Alexandra
authors.initials: A
authors.lastname: Salvador
authors.forename: J Mark
authors.initials: JM
authors.lastname: Carter
mesh_terms: D000906:Antibodies / Q000737:chemistry*; D057205:Automation, Laboratory; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D019453:Escherichia coli O157 / Q000254:growth & development / Q000472:pathogenicity; D057166:High-Throughput Screening Assays / Q000295:instrumentation*; D008070:Lipopolysaccharides / Q000032:analysis*; D008863:Microspheres; D010799:Phycoerythrin / Q000737:chemistry; D015203:Reproducibility of Results; D022622:Shiga Toxin 1 / Q000032:analysis*; D022641:Shiga Toxin 2 / Q000032:analysis*; D013535:Suspensions / Q000737:chemistry
chemical_list: D000906:Antibodies; D008070:Lipopolysaccharides; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2; D013535:Suspensions; D010799:Phycoerythrin