Studies on multivalent interactions of quantum dots-protein self-assemble using fluorescence coupled capillary electrophoresis

摘要

Nanoparticle-biomolecules self-assembly is the key to the understanding of biomolecular coating of nanoparticle. However, the self-assembly of biomolecules with nanoparticles is still under-exploited because of the lack of an efficient method to detect the subtle changes in the surface of nanoparticles. In this study, we utilized fluorescence coupled capillary electrophoresis (CE-FL) to probe the binding interaction between a multivalent ligand (dBSA, denatured bovine serum albumin which contains multiple thiol groups) and CdSe/ZnS quantum dots (QDs, 5 nm in diameter). The yield of QDs-dBSA complex increased with increasing molar ratio of dBSA to QDs, which plateaued at a ratio of 8:1. Besides, QDs-dBSA complex showed good stability due to the multivalent interaction, revealing that dBSA is a superior ligand for QDs. The self-assembly kinetics of QDs with dBSA manifested a bi-phasic kinetics with a linear initial stage followed by a saturating stage. This work revealed for the first time that there exist two types of binding sites on the surface of QDs for dBSA: one type termed "high priority" binding sites, which preferentially bind to the protein, whereas the "low priority" sites are occupied only after the first-type binding sites are fully bound. Our work thereby represents the first example of systematic investigation on the details of the metal-affinity driven selfassembly between QDs and dBSA utilizing the highresolution CE-FL. It also expanded the application of CE-FL in the study of nanoparticle-biomolecule interaction and kinetics analysis.