Surface Analysis of Nanocomplexes by X-ray Photoelectron Spectroscopy (XPS)

Self-assembled nanocomplexes composed of individual molecules that spontaneously connect via noncovalent interactions have recently emerged as versatile alternatives to conventional controlled drug delivery systems because of their unique bioinspired properties (responsiveness, dynamics, etc.). Characterization of such nanocomplexes typically includes their size distribution, surface charge, morphology, drug entrapment efficiency, and verification of the coexistence of labeled components within the nanocomplexes using a colocalization study. Less common is the direct examination of the molecular interactions between the different components in the coassembled nanocomplex, especially in nanocomplexes composed of hygroscopic components, because convenient methods are still lacking. Here, we present a detailed experimental protocol for determining the surface composition and the chemical bonds by X-ray photoelectron spectroscopy (XPS) after drying the deposit hygroscopic sample overnight under UHV. We applied this method to investigate the surface chemistry of binary Ca²⁺-siRNA nanocomplexes and ternary nanocomplexes of hyaluronan-sulfate (HAS)-Ca²⁺-siRNA, deposited on a wafer. Notably, we showed that the protocol can be implemented to study the surface composition and interactions of the deposited nanocomplexes with a traditional XPS instrument, and it requires only a relatively small amount of the nanocomplex suspension.