The Competitive Dynamic Binding of Some Blood Proteins Adsorbed on Gold Nanoparticles

Abstract

Nanoparticle (NP) surfaces are modified immediately by the adsorption of proteins when injected into human blood, leading to the formation of a protein corona. The protein‐coated NPs may be recognized by living cells. Furthermore, the adsorption of serum proteins is a continuous competitive dynamic process that is the key to exploring the bioapplication and biosafety of NPs. In this study, the competitive dynamic adsorption of some serum proteins on gold nanoparticles (AuNPs) is investigated by fluorescence emission, dynamic light scattering, and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. Serum proteins with different AuNPs binding affinities are used to address the competitive dynamic process of protein‐AuNP interactions in vitro. The results show that more abundant serum proteins, such as human serum albumin, adsorb on AuNPs first, and then the higher binding affinity and lower concentration serum proteins, such as fibrinogen (FIB), replace the abundant and lower binding affinity serum proteins. However, the lower binding affinity serum proteins, such as hemoglobin, do not replace the higher binding affinity proteins from the protein‐AuNP conjugates. During the dynamic exchange process, the larger the binding affinities difference between two proteins, the faster the exchange rate. This dynamic exchange process usually takes longer in inner protein‐AuNP conjugates (hard corona) than the external surface of protein‐AuNP conjugates (soft corona).

Conformational-transited protein corona regulated cell-membrane penetration and induced cytotoxicity of ultrasmall Au nanoparticles

“…The mixture was maintained at a pH of 8.5 to ensure monolabeling. 41 The protein conjugates were separated from the mixture by passing through a Sephadex G-25 column and eluted with 10 Â 10 À3 M phosphate buffer solution (PBS, pH 7.2). The effluent was collected from 2.5 to 6 mL.…”

Solution NMR of Nanoparticles in Serum: Protein Competition Influences Binding Thermodynamics and Kinetics

“…These results reflect the fact that DLS is highly sensitive to the change in particle size resulting from protein adsorption. SDS-PAGE analysis of serum-AuNP adsorption by others demonstrates that ∼90% of adsorbed proteins are bovine serum albumin (BSA, Figure S1, Supplementary Material) (Zhang et al, 2019). It is therefore hypothesized that the majority of proteins in the FBS-AuNP corona are BSA proteins, which have a protein size ∼10 times of GB3 (6.2 kDa).…”

“…These approaches are often able to differentiate many proteins, but they are often unable to study nanoparticle binding in situ. Specifically, mass spectrometric and electrophoretic techniques are widely used to quantify multiple proteins attached in the protein corona (Monopoli et al, 2011;Mo et al, 2018;Zhang et al, 2019;Han et al, 2020;Madathiparambil Visalakshan et al, 2020;Moon et al, 2020;Pinals et al, 2020;Liessi et al, 2021) even at the detail of individual protein residues (Pustulka et al, 2020). However, these methods require extensive purification of the protein-nanoparticle complexes and complete displacement of bound proteins from the nanoparticle surface for analysis.…”

“…Multiple techniques exist to monitor thermodynamics and kinetics of binding for purified proteins on a uniform population of nanoparticles; however, this type of reductionist approach may not always work (Lacerda et al, 2010;Lee et al, 2020). Specifically, multiple studies have found that the binding affinity of an individual protein fails to predict its composition in the final protein corona formed in a mixture (Monopoli et al, 2011;Zhang et al, 2019). Indeed, the mixture itself will have a significant influence on the protein(s) of interest (Mahmoudi et al, 2011).…”

Probing protein dissociation from gold nanoparticles and the influence of temperature from the protein corona formation mechanism

“…Protein corona formation is a complex process and is kinetically divided into two phases. 17,18 In phase one, proteins are absorbed on the NP surface in seconds or minutes by fast electrostatic or hydrophobic forces. For example, more than 300 plasma protein species rapidly adsorb onto AuNPs within 0.5 min.…”