title: Development of a homogeneous immunoassay system using protein A fusion fragmented Renilla luciferase.
abstract: A homogeneous immunoassay system was developed using fragmented Renilla luciferase (Rluc). The B domain of protein A was fused to two Rluc fragments. When complexes between an antibody and fragmented Rluc fusion proteins bind to target molecules, the Rluc fragments come into close proximity and the luminescence activity of fragmented Rluc is restored by complementation. As proof-of-principle, this fragmented Rluc system was used to detect E. coli homogeneously using an anti-E. coli antibody.
authors.affiliation: Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, 226-8501, Japan.
authors.forename: Masayasu
authors.initials: M
authors.lastname: Mie
authors.forename: Ngo Phan Bich
authors.initials: NP
authors.lastname: Thuy
authors.forename: Eiry
authors.initials: E
authors.lastname: Kobatake
mesh_terms: D000907:Antibodies, Bacterial / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D007118:Immunoassay / Q000379:methods*; D049410:Luciferases, Renilla / Q000737:chemistry*; D010446:Peptide Fragments / Q000235:genetics*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D013205:Staphylococcal Protein A / Q000737:chemistry*
chemical_list: D000907:Antibodies, Bacterial; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D049410:Luciferases, Renilla