title: Phage anti-immune complex assay: general strategy for noncompetitive immunodetection of small molecules.
abstract: Due to their size, small molecules cannot be simultaneously bound by two antibodies, precluding their detection by noncompetitive two-site immunoassays, which are superior to competitive ones in terms of sensitivity, kinetics, and working range. This has prompted the development of anti-immune complex antibodies, but these are difficult to produce, and often exhibit high cross-reactivity with the unliganded primary antibody. This work demonstrates that anti-immune complex antibodies can be substituted by phage particles isolated from phage display peptide libraries. Phages bearing specific small peptide loops allowed to focus the recognition to changes in the binding area of the immune complex. The concept was tested using environmental and drug analytes; with improved sensitivity and ready adaptation into on-site formats. Peptides specific for different immune complexes can be isolated from different peptide libraries in a simple and systematic fashion allowing the rapid development of noncompetitive assays for small molecules.
authors.affiliation: Cátedra de Inmunología, Facultad de Química, Instituto de Higiene, UDELAR, Montevideo, Uruguay 11600.
authors.forename: A
authors.initials: A
authors.lastname: González-Techera
authors.forename: L
authors.initials: L
authors.lastname: Vanrell
authors.forename: J A
authors.initials: JA
authors.lastname: Last
authors.forename: B D
authors.initials: BD
authors.lastname: Hammock
authors.forename: G
authors.initials: G
authors.lastname: González-Sapienza
mesh_terms: D000936:Antigen-Antibody Complex / Q000032:analysis* / Q000276:immunology*; D001667:Binding, Competitive; D007118:Immunoassay / Q000379:methods*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D012680:Sensitivity and Specificity
chemical_list: D000936:Antigen-Antibody Complex; D019151:Peptide Library; D010455:Peptides