title: Graph-based deconvolution analysis of multiplex sandwich microarray immunoassays: applications for environmental monitoring.
abstract: The sandwich microarray immunoassay (SMI) is a powerful technique for the analysis and characterization of environmental samples, from the identification of microorganisms to specific bioanalytes. As the number of antibodies increases, however, unspecific binding and cross-reactivity can become a problem. To cope with such difficulties, we present here the concept of antibody graph associated to a sandwich antibody microarray. Antibody graphs give valuable information about the antibody cross-reactivity network and all the players involved in the sandwich format: capturing and tracer antibodies, the antigenic sample and the degree of cross-reactivity between antibodies. Making use of the information contained in the antibody graph, we have developed a deconvolution method that disentangles the antibody cross-reactivity events and gives qualitative information about the composition of the experimental sample under study. We have validated the method by using a 66 antibody-containing microarray to describe known antigenic mixtures as well as natural environmental samples characterized by 16S-RNA gene phylogenetic analysis. The application of our antibody graph and deconvolution method allowed us to discriminate between true specific antigen-antibody reactions and spurious signals on a microarray designed for environmental monitoring.
authors.affiliation: Department of Molecular Evolution, Centro de Astrobiología (INTA-CSIC), Madrid, Spain. rivasmla@inta.es
authors.forename: Luis A
authors.initials: LA
authors.lastname: Rivas
authors.forename: Jacobo
authors.initials: J
authors.lastname: Aguirre
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Blanco
authors.forename: Elena
authors.initials: E
authors.lastname: González-Toril
authors.forename: Víctor
authors.initials: V
authors.lastname: Parro
mesh_terms: D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D003429:Cross Reactions; D004783:Environmental Microbiology; D004784:Environmental Monitoring / Q000379:methods*; D007118:Immunoassay / Q000379:methods; D010802:Phylogeny; D040081:Protein Array Analysis; D012336:RNA, Ribosomal, 16S
chemical_list: D000906:Antibodies; D012336:RNA, Ribosomal, 16S