Fabrication of Orange-Emitting Organic Nanoparticle-Protamine Conjugate: Fluorimetric Sensor of Heparin.

Abstract

Among the diverse sensing techniques, fluorimetric detection dominates over the other methods because of its rapid signaling, high selectivity and sensitivity, and operational simplicity. This present article delineates fabrication of a fluorescent organic nanoparticle-protamine (FONP-Pro) conjugate for selective and sensitive detection of heparin simply by exploitation of the aggregation-induced emission (AIE) property of the FONPs. Naphthalene diimide-based bola-type amphiphilic molecules (NDI-1) comprise a naphthyl residue and a 3-aminopyridyl unit at both terminals, forming organic nanoparticles in a dimethyl sulfoxide-water binary solvent mixture, and exhibited AIE through excimer formation. The presence of naphthyl residue in the molecular backbone facilitates the intramolecular charge transfer to generate orange-emitting (λem = 594 nm) AIE-luminogen (AIE-gen). The aminopyridine residues within NDI-1 induced negative surface charge on NDI-1 FONPs, which facilitated interaction with positively charged protamine (Pro) to construct FONP-Pro conjugates. Formation of this NDI-1 FONP-Pro conjugate through the interaction between Pro and FONP drastically reduced the orange emission intensity (fluorescence off) of the AIE-gens. Interestingly, addition of heparin to this FONP-Pro conjugate turned on the fluorescence signal of FONPs through unwinding of the Pro from the FONP surface because of a strong binding affinity between heparin and Pro. Formation of the FONP-Pro conjugate and fluorimetric sensing of heparin was investigated by monitoring the change in emission behavior of NDI-1 FONPs. Also, the heparin-sensing was found to be highly selective against many other biomolecules including proteins, enzymes, and DNA. Hence, a selective and efficient heparin sensor (FONP-Pro) was developed having a limit of detection of 12 nM simply by utilizing the fluorescence "turn-off" and "turn-on" mechanism of NDI-1 FONP.