Bioluminescent Antibodies through Photoconjugation of Protein G-Luciferase Fusion Proteins.

ABSTRACT: Bioluminescent antibodies represent attractive detection agents in both bioanalytical assays and imaging. Currently, their preparation relies on genetic fusion of luciferases to antibodies or nonspecific chemical conjugation strategies. Here, we report a generic method to generate well-defined covalent antibody-luciferase conjugates starting from commercially available monoclonal antibodies. Our approach uses fusion proteins consisting of the bright blue light-emitting luciferase NanoLuc (NL) and an Fc-binding protein domain (Gx) that can be photo-cross-linked to the antibody using UV light illumination. Green and red color variants were constructed by tight fusion of the NanoLuc with a green fluorescent acceptor domain and introduction of Cy3, respectively. To increase the already bright

SUBMITTER: Wouters SFA 

PROVIDER: S-EPMC7086395 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature