A Universal and Programmable Platform based on Fluorescent Peptide‐Conjugated Probes for Detection of Proteins in Organelles of Living Cells

Realizing protein analysis in organelles of living cells is of great significance for developing diagnostic and therapeutic methods of diseases. Fluorescent‐labeled antibodies with well imaging performance and high affinity are classical biochemical tools for protein analysis, while due to the inability to effectively enter into cells, not to mention organelles and the uncontrollable reaction sites that might cause antibodies inactivation when chemically modification, they are hard to apply to living cells. Inspired by the structure of fluorescent‐labeled antibodies, we designed as a universal detection platform that was based on the peptide‐conjugated probes (PCPs) and consisted of three parts: a) a rotor type fluorescent molecular scaffold for conjugation and signal output; b) the cell penetration protein recognition unit; c) the subcellular organelle targeting unit. In living cells, PCPs could firstly localize at organelles and then proceed protein specific recognition, thus jointly leading to the restriction of twisted intramolecular charge transfer and activation of fluorescence signal. As a proof‐of‐concept, six different proteins in three typical intracellular organelles could be detected by our platform through simply replacing the recognition sequence of proteins and matching organelle targeting units. The position and intensity of fluorescence signals demonstrated specificity of PCPs and universality of the platform. A universal detection platform based on the peptide‐conjugated probes (PCPs, name as TP‐TPE‐TO) by integrating protein recognition units, signal units, and particularly introducing organelle targeting units. The universal detection platform can achieve six protein analysis in three different organelles through simple transformation of recognition units and targeting units.