title: Combination of UV-vis spectroscopy and chemometrics to understand protein-nanomaterial conjugate: a case study on human serum albumin and gold nanoparticles.
abstract: Study of the interactions between proteins and nanomaterials is of great importance for understanding of protein nanoconjugate. In this work, we choose human serum albumin (HSA) and citrate-capped gold nanoparticles (AuNPs) as a model of protein and nanomaterial, and combine UV-vis spectroscopy with multivariate curve resolution by an alternating least squares (MCR-ALS) algorithm to present a new and efficient method for comparatively comprehensive study of evolution of protein nanoconjugate. UV-vis spectroscopy coupled with MCR-ALS allows qualitative and quantitative extraction of the distribution diagrams, spectra and kinetic profiles of absorbing pure species (AuNPs and AuNPs-HSA conjugate are herein identified) and undetectable species (HSA) from spectral data. The response profiles recovered are converted into the desired thermodynamic, kinetic and structural parameters describing the protein nanoconjugate evolution. Analysis of these parameters for the system gives evidence that HSA molecules are very likely to be attached to AuNPs surface predominantly as a flat monolayer to form a stable AuNPs-HSA conjugate with a core-shell structure, and the binding process takes place mainly through electrostatic and hydrogen-bond interactions between the positively amino acid residues of HSA and the negatively carboxyl group of citrate on AuNPs surface. The results obtained are verified by transmission electron microscopy, zeta potential, circular dichroism spectroscopy and Fourier transform infrared spectroscopy, showing the potential of UV-vis spectroscopy for study of evolution of protein nanoconjugate. In parallel, concentration evolutions of pure species resolved by MCR-ALS are used to construct a sensitive spectroscopic biosensor for HSA with a linear range from 1.8 nM to 28.1 nM and a detection limit of 0.8 nM.
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China; Department of Chemistry, Nanchang University, Nanchang 330031, Jiangxi, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China; Department of Chemistry, Nanchang University, Nanchang 330031, Jiangxi, China. Electronic address: ynni@ncu.edu.cn.
authors.forename: Yongnian
authors.initials: Y
authors.lastname: Ni
mesh_terms: D006046:Gold / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D053768:Metal Nanoparticles; D046529:Microscopy, Electron, Transmission; D049329:Nanostructures; D012709:Serum Albumin / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D013816:Thermodynamics
chemical_list: D012709:Serum Albumin; D006046:Gold
keywords: Biosensor; Chemometrics; Gold nanoparticles; Protein nanoconjugate; Thermodynamics; UV–vis spectroscopy