Evaluation of Dibenzocyclooctyne and Bicyclononyne Click Reaction on Azido?Functionalized Antifouling Polymer Brushes via Microspotting

Strain-promoted alkyne-azide cycloaddition (SPAAC) has become an indispensable tool in bioorthogonal conjugation and surface immobilization. While numerous studies have focused on enhancing the reactivity of cyclooctynes, a facile method to evaluate binding efficiency for cyclooctyne-azide-based immobilization without any sophisticated facilities is still missing. In present work, different derivatives dibenzocyclooctyne/bicyclononyne (DBCO/BCN) linked either fluorophore or biotin-moiety are patterned ultra-low fouling polymer brushes, which can avoid unspecific protein contamination prior blocking steps. The brushes composed antifouling bottom block azide-terminated top block. assessment conducted ordered arrays spotted by microchannel cantilever spotting (?CS) with normal fluorescent microscope. Both cyclooctynes demonstrate reliable performance azide-bearing diblock via ?CS, but DBCO shows higher density molecular according assays. This work provides reference choosing appropriate cyclooctyne couple azides be useful design biosensors bio-platforms analyte detection, cell capture, other biological applications.