Assembly and Detachment of Hyaluronic Acid on a Protein-Conjugated Gold Nanoparticle

The assembly-disassembly of hyaluronic acid (HA) with a bovine serum albumin-conjugated gold nanoparticle (BSA-AuNP) was demonstrated using a gas-phase electrophoresis approach, electrospray-differential mobility analysis (ES-DMA). Physical sizes, number and mass concentrations, and degrees of aggregation of HA, BSA, and AuNP were successfully quantified using ES-DMA hyphenated with inductively coupled plasma mass spectrometry. Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy was employed complementarily for an orthogonal characterization of the assembly of HA with BSA-AuNP and the subsequent HA detachment. The results show that the surface packing density of HA on BSA-AuNP was proportional to the concentration of HA (C-HA) when C-HA <= 5 X 10(-3) mu mol/L, and the equilibrium binding constant of HA on BSA-AuNP was identified as approximate to 4 X 10(5) L/mol at pH 3. The pH-sensitive and enzyme-induced detachments of HA from BSA-AuNP were both successfully characterized using ES-DMA and ATR-FTIR. In the absence of enzymatic catalysis, the rate constant of HA detachment (k) was shown to increase by at least 3.7 times on adjusting the environmental acidity from pH 3 to pH 7. A significant enzyme-induced HA detachment was identified at pH 7, showing a remarkable increase of k by at least two times in the presence of an enzyme. This work provides a proof of concept for assembly of HA-based hybrid colloidal nanomaterials through the tuning of surface chemistry in the aqueous phase with the ability of in situ quantitative characterization, which has shown promise for the development of a variety of HA-derivative biomedical applications (e.g., drug delivery).