摘要：SummaryPhotochemical transformations enable exquisite spatiotemporal control over biochemical processes; however, methods for reliable manipulations of biomolecules tagged with biocompatible photo-sensitive reporters are lacking. Here we created a high-affinity binder specific to a photolytically removable caging group. We utilized chemical modification or genetically encoded incorporation of noncanonical amino acids to produce proteins with photocaged cysteine or selenocysteine residues, which were used for raising a high-affinity monoclonal antibody against a small photoremovable tag, 4,5-dimethoxy-2-nitrobenzyl (DMNB) group. Employing the produced photocage-selective binder, we demonstrate selective detection and immunoprecipitation of a variety of DMNB-caged target proteins in complex biological mixtures. This combined orthogonal strategy permits photocage-selective capture and light-controlled traceless release of target proteins for a myriad of applications in nanoscale assays.Graphical AbstractDisplay OmittedHighlights•The first high-affinity monoclonal antibody specific for a popular photocaging group•A new tool for selective detection of DMNB-tagged proteins in complex mixtures•Enables non-covalent capture of native proteins with surface-exposed DMNB groups•Orthogonal protein manipulation by photocage-selective capture and photolytic releaseBiochemistry; Biochemistry Methods; Biotechnology
