title: Detection of small molecule diagnostic markers with phage-based open-sandwich immuno-PCR.
abstract: Open sandwich immunoassay (OS-IA) utilizes antigen-dependent stabilization of antibody variable region to quantify various antigens, enabling noncompetitive detection of small molecules with a broad working range. To further improve its detection sensitivity, here we employed phage-based immuno-PCR approach, wherein OS-IA and quantitative PCR methodologies were combined with the use of immobilized V(L) fusion protein and filamentous phages displaying V(H) fragment, whose DNA was extracted for PCR amplification. This approach significantly enhanced the assay sensitivity for small molecule antigens osteocalcin (BGP) peptide and 17beta-estradiol.
authors.affiliation: Department of Chemistry and Biotechnology, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8656, Japan.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Dong
authors.forename: Sharif
authors.initials: S
authors.lastname: Hasan
authors.forename: Yuuichiro
authors.initials: Y
authors.lastname: Fujioka
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ueda
mesh_terms: D000941:Antigens / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D001435:Bacteriophages / Q000235:genetics / Q000276:immunology*; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000032:analysis / Q000235:genetics / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007135:Immunoglobulin Variable Region / Q000737:chemistry* / Q000276:immunology; D057230:Limit of Detection; D015675:Osteocalcin / Q000032:analysis / Q000235:genetics / Q000276:immunology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D000941:Antigens; D007135:Immunoglobulin Variable Region; D015675:Osteocalcin; D004958:Estradiol