Selection of optical biosensors from chemisynthetic antibody libraries.
We describe a method for creating antibodies with a fluorescent reporter integrated into the antigen-binding site. A reporter molecule was chemically linked to a hypervariable loop of an antibody repertoire displayed on phage, and this repertoire was selected for antigen binding. In one selected antibody, the fluorescence of the probe responded quantitatively to antigen binding. The method may have application for the engineering of homogeneous immunoassays.
Jespers
Laurent
L
Division for Protein and Nucleic Acid Chemistry, Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.
Bonnert
Timothy P
TP
Winter
Greg
G
D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry* / Q000235:genetics; D000937:Antigen-Antibody Reactions; D001666:Binding Sites, Antibody; D015374:Biosensing Techniques / Q000379:methods*; D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000379:methods; D066298:In Vitro Techniques; D008958:Models, Molecular; D055095:Optics and Photonics; D019151:Peptide Library; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
D000906:Antibodies; D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D019151:Peptide Library; D011994:Recombinant Proteins