Rational Synthesis of Imine-Linked Fluorescent Covalent Organic Frameworks with Different pKa for pH Sensing In Vitro and In Vivo.

Precise modulation of pH in living cells plays a vital role in the study of many diseases, such as cancer and rheumatoid arthritis. Here, a series of imine-linked covalent organic frameworks (COFs) were rationally designed and developed for pH sensing in tumor cells and zebrafish. Four monomers were chosen to synthesize COFs (COF1-COF4) with different pKa by a simple orthogonal combination through condensation reaction. The as-obtained COFs exhibited a sensitive pH-dependent fluorescence response compared to their building blocks. Among them, COF2 possessed a high crystallinity, excellent fluorescence, and suitable pKa for biosensing. For bioimaging applications, COF2 was modified with poly-d-lysine (PDL) to improve its biocompatibility and endocytosis efficiency. After that, PDL-modified COF2 (PDL@COF2) was used as a novel fluorescence probe with a superior linear pH response over the range from 5.0 to 8.0 due to its fully reversible protonation and deprotonation. The fluorescent PDL@COF2 was further employed as a good candidate for pH imaging in tumor cells and zebrafish. The as-constructed environment-sensitive fluorescent COFs have greatly expanded the applications of COFs in the biological area.