Rapid conjugation of nanoparticles, proteins and siRNAs to microbubbles by strain-promoted click chemistry for ultrasound imaging and drug delivery

A new strategy using catalyst-free strain-promoted alkyne–azide cycloaddition (SPAAC) “click” chemistry for the ligation of anti-cancer drug-loaded nanoparticles, functionalized proteins, and siRNA conjugated micelles to microbubbles (MB) was established. The results showed fast ligation within 5 min without sacrificing microbubble size and density. The ultrasound test showed good imaging abilities of the microbubbles after functionalization. This microbubble–therapeutic SPAAC “click” conjugation developed in the current study involves no toxic catalyst or initiator, has ultra-fast reaction speed, and is versatile for the ligation of various anti-cancer or therapeutic agents to microbubbles. These advantages render the SPAAC click strategy promising for broad applications in ultrasound-guided imaging and therapeutic delivery.