Citrate-coated AuNPs Research Articles

Efficiency of three dimension Au–MnO2 nanostructure as visible adsorption based detector of histamine sensing

High intakes of histamine are poisonous. Hence, determining histamine levels in fisheries and processed products is essential to food quality. Several colorimetric sensors based on gold nanoparticles have shown exemplary performance in detecting the presence of histamine in samples. The modified AuNPs by MnO2 were used as a reagent in histamine analysis by UV–Vis spectrophotometry. Using a seed-mediated growth method, modified Au–MnO2 nanostructures were synthesized from citrate-coated AuNPs and KMnO4. The Au–MnO2 nanostructure synthesis process produced a yellow-green solution, which produced a crystalline solid with a particle size <100 nm by evaporation. These nanoparticles comprise AuNPs and MnO2 components, which still perform their original properties. Application of Au–MnO2 nanostructure in histamine spectrophotometry has values in sensitivity of 0.0344 ppm, detection limit of 0.185 ppm, quantification limit of 0.618 ppm, and accuracy of 95–101 %. Based on analytical results, the effectiveness of synthesizing Au–MnO2 nanostructure to form complexes with histamine content was low, and it was due to the weak binding between the imidazole group of histamine and Au–MnO2 nanostructures. Therefore, further investigation on the composition of the Mn metal or the selection of other metals is needed to use Au–MnO2 as candidate histamine biosensors.

Adsorption and Molecular Display of a Redox-Active Protein on Gold Nanoparticle Surfaces.

Engineered gold nanoparticles (AuNPs) have great potential in many applications due to their tunable optical properties, facile synthesis, and surface functionalization via thiol chemistry. When exposed to a biological environment, NPs are coated with a protein corona that can alter the NPs' biological identity but can also affect the proteins' structures and functions. Protein disulfide isomerase (PDI) is an abundant protein responsible for the disulfide formation and isomerization that contribute to overall cell redox homeostasis and signaling. Given that AuNPs are widely employed in nanomedicine and PDI plays a functional role in various diseases, the interactions between oxidized (oPDI) and reduced (rPDI) with 50 nm citrate-coated AuNPs (AuNPs) are examined in this study using various techniques. Upon incubation, PDI adsorbs to the AuNP surface, which leads to a reduction in its enzymatic activity despite limited changes in secondary structures. Partial enzymatic digestion followed by mass spectrometry analysis shows that orientation of PDI on the NP surface is dependent on both its oxidation state and the PDI:AuNP incubation ratios.

Controlling plasmonic suprastructures through self-assembly of gold nanoparticles with hybrid copolymer-lipid vesicles

Hybrid lipid membranes incorporating amphiphilic copolymers have gained significant attention due to their potential applications in various fields, including drug delivery and sensing. By combining the properties of copolymers and lipid membranes, such as enhanced chemical tunability and stability, environmental responsiveness, and multidomain nature, novel membrane architectures have been proposed. In this study, we investigated the potentialities of hybrid membranes made of two distinct components: the rigid fully saturated phospholipid 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and the soft copolymer poly(butadiene-b-ethyleneoxide) (PBD-b-PEO). The objective was to explore the interaction of citrate-coated gold nanoparticles (AuNPs) and the hybrid membrane, aiming at constructing AuNPs-hybrid vesicles suprastructures with controlled and adjustable plasmonic properties. A series of experimental techniques were employed to investigate hybrid free-standing and supported membranes. The results revealed that the incorporation of the copolymer into the lipid membrane promotes AuNPs clustering, demonstrating a distinctive aggregative phenomenon of citrate-coated AuNPs on multidomain membranes. Importantly, we show that the size and morphology of AuNPs clusters can be precisely controlled in non-homogeneous membranes, enabling the formation of hybrid suprastructures with controlled patch properties. These results highlight the potential of lipid-copolymer hybrid membranes for designing functional materials with tailored plasmonic properties, with potential applications in nanomedicine and sensing.

Goldilocks Energy Minimum: Peptide-Based Reversible Aggregation and Biosensing.

Colorimetric biosensors based on gold nanoparticle (AuNP) aggregation are often challenged by matrix interference in biofluids, poor specificity, and limited utility with clinical samples. Here, we propose a peptide-driven nanoscale disassembly approach, where AuNP aggregates induced by electrostatic attractions are dissociated in response to proteolytic cleavage. Initially, citrate-coated AuNPs were assembled via a short cationic peptide (RRK) and characterized by experiments and simulations. The dissociation peptides were then used to reversibly dissociate the AuNP aggregates as a function of target protease detection, i.e., main protease (Mpro), a biomarker for severe acute respiratory syndrome coronavirus 2. The dissociation propensity depends on peptide length, hydrophilicity, charge, and ligand architecture. Finally, our dissociation strategy provides a rapid and distinct optical signal through Mpro cleavage with a detection limit of 12.3 nM in saliva. Our dissociation peptide effectively dissociates plasmonic assemblies in diverse matrices including 100% human saliva, urine, plasma, and seawater, as well as other types of plasmonic nanoparticles such as silver. Our peptide-enabled dissociation platform provides a simple, matrix-insensitive, and versatile method for protease sensing.

Lectin-Triggered Aggregation of Glyco-Gold Nanoprobes for Activity-based Sensing of Hydrogen Peroxide by the Naked Eye.

The purpose of this study was to develop a colorimetric assay for detecting hydrogen peroxide (H2 O2 ) through a combination of using an aryl boronate (AB) derivative and gold nanoparticles (AuNPs). The unique optical property of AuNPs is applied to design a detection probe. The aggregation of AuNPs could be directly observed as a color change by the naked eye. A mannoside-boronate-sulfide (MBS) ligand was designed that contains an arylboronate (AB), a mannoside, and a thiol group. The thiol group bonds covalently with the surface of AuNPs to obtain MBS@AuNPs. The mannoside moiety recognizes concanavalin A (Con A), a lectin with four carbohydrate recognition sites that can specifically recognize the non-reducing end of an α-D-mannoside or α-D-glucoside structure. The AB structure on MBS first reacts with H2 O2 and then inserts an oxygen atom in the B-H bond, which triggers intramolecular electron rearrangement to cleave the covalent bond, resulting in a MBSt mixture. The MBS or MBSt is then modified to citrate-coated AuNPs (c-AuNPs) to have MBS@AuNPs or MBSt@AuNPs. When the MBS@AuNPs are incubated with Con A, the Con A recognizes multiple mannosides on the surface of the MBS@AuNPs. Subsequently, the MBS@AuNPs aggregate and the solution's color changes from red to purple, but this color change does not occur in the case of MBSt@AuNPs. The phenomenon can be observed by the naked eye.

Gold Nanoparticles Dissolve Extracellularly in the Presence of Human Macrophages.

IntroductionGold nanoparticles (AuNPs) have the potential to be used in various biomedical applications, partly due to the inertness and stability of gold. Upon intravenous injection, the NPs interact with the mononuclear phagocyte system, first with monocytes in the blood and then with macrophages in tissue. The NP–macrophage interaction will likely affect the stability of the AuNPs, but this is seldom analyzed. This study aimed to elucidate the role of macrophages in the biodissolution of AuNPs and underlying mechanisms.MethodsWith an in vitro dissolution assay, we used inductively coupled plasma mass spectrometry to quantitatively compare the dissolution of 5 and 20 nm AuNPs coated with citrate or PEG in cell medium alone or in the presence of THP1-derived macrophages at 24 hours. In addition, we analyzed the cell dose, compared extra- and intracellular dissolution, and explored the possible role of reactive nitrogen species.ResultsThe results showed a higher cellular dose of the citrate-coated AuNPs, but dissolution was mainly evident for those sized 5 nm, irrespective of coating. The macrophages clearly assisted the dissolution, which was approximately fivefold higher in the presence of macrophages. The dissolution, however, appeared to take place mainly extracellularly. Acellular experiments demonstrated that peroxynitrite can initiate oxidation of gold, but a ligand is required to keep the gold ions in solution.ConclusionThis study suggests extracellular dissolution of AuNPs in the presence of macrophages, likely with the contribution of the release of reactive nitrogen species, and provides new insight into the fate of AuNPs in the body.

Subchronic oral toxicity evaluation of gold nanoparticles in male and female mice

Gold nanoparticles (AuNPs) are biocompatible nanomaterials with potential application in the food industry. The safety of AuNPs oral consumption remains inconclusive, and information on possible long-term toxicity is limited. The current study aimed to evaluate the subchronic oral toxicity of AuNPs in male and female Institute of Cancer Research (ICR) mice. Citrate-coated spherical AuNPs with 53 nm diameters were prepared and orally administered to the mice. No mortality or clinical abnormalities were observed following daily administration of AuNPs at the dosages of 0.2, 2, and 20 mg/kg for 90 days. There was no significant difference in body weight or the relative organs' weights between the control and AuNPs-treated mice. No gross abnormalities or histopathological changes were observed except that the male mice treated with high dose (20 mg/kg AuNPs) showed minor infiltration in the kidneys, and female mice showed a reduced A/G ratio and elevated platelet indices. Overall, the 90-day long-term oral consumption of AuNPs did not cause significant toxicity in mice.

Transient Vasodilation in Mouse 4T1 Tumors after Intragastric and Intravenous Administration of Gold Nanoparticles.

Gold nanoparticles (AuNPs) are foreseen as a promising tool in nanomedicine, both as drug carriers and radiosensitizers. They have been also proposed as a potential anticancer drug due to the anti-angiogenic effect in tumor tissue. In this work we investigated the effect of citrate-coated AuNPs of nominal diameter 20 nm on the growth and metastatic potential of 4T1 cells originated from a mouse mammary gland tumor inoculated into the mammary fat pad of Balb/ccmdb mice. To evaluate whether AuNPs can prevent the tumor growth, one group of inoculated mice was intragastrically (i.g.) administered with 1 mg/kg of AuNPs daily from day 1 to day 14 after cancer cell implantation. To evaluate whether AuNPs can attenuate the tumor growth, the second group was intravenously (i.v.) administered with 1 or 5 mg/kg of AuNPs, twice on day 5 and day 14 after inoculation. We did not observe any anticancer activity of i.v. nor i.g. administered AuNPs, as they did not affect neither the primary tumor growth rate nor the number of lung metastases. Unexpectedly, both AuNP treatment regimens caused a marked vasodilating effect in the tumor tissue. As no change of potential angiogenic genes (Fgf2, Vegfa) nor inducible nitric oxygenase (Nos2) was observed, we proposed that the vasodilation was caused by AuNP-dependent decomposition of nitrosothiols and direct release of nitric oxide in the tumor tissue.

Biogenic Synthesis of Gold Nanoparticles by Bacteria and Utilization of the Chemical Fabricated for Diagnostic Performance of Viral Hepatitis C Virus-NS4

Extracellular biosynthesis of gold nanoparticles (AuNPs) becomes a significant approach to NPs preparation. Development in medical diagnostics focuses on the nanomaterials, particularly AuNPs. This study focuses on the biogenic synthesis of AuNPs and, additionally, the use of the chemicals made-up in medical detection of hepatitis C Virus-NS4 (HCV-NS4). Cell-free extracts (CFE's) and cell biomass (CBM) of E. coli ATCC 8739, B. subtilis ATCC 6633, St. thermophilus ESH1 and L. acidophilus DSMZ 20079T mixed with Tetrachloroauric acid (HAuCl4) at 1:1(v/v) ratio, kept at distinctive temperatures and pHs for time intervals, and observed for AuNPs biogenesis. The chemically made-up AuNPs, 5'- hexanethiol, and the critical 20-base target oligonucleotide mixed. Single-stranded DNA (ssDNA) adsorbate on citrate-coated AuNPs used for the detection of the HCV-NS4 target oligonucleotide sequence. This study has advanced fast biogenic synthesis of stable AuNPs from 10-3M HAuCl4 utilizing the bacteria mentioned above at 37 oC, pH 7, and within fifteen minutes. UV spectrum at 520 nm, XRD, and TEM assured the development of AuNPs. Incorporated created up AuNPs recorded broad uptake bands and obtained the color modification from crimson to blue. The results directed to the biogenic synthesis of AuNPs and the attainable utilization of made-up AuNPs to seek out HCV-NS4.

Shedding light on membrane-templated clustering of gold nanoparticles

The use of inorganic nanoparticles in biomedical and biotechnological applications requires a molecular-level understanding of interactions at nano-bio interfaces, such as cell membranes. Several recent reports have shown that gold nanoparticles (AuNP), in the presence of fluid lipid bilayers, aggregate at the lipid/aqueous interface, but the precise origin of this phenomenon is still not fully understood. Here, by challenging synthetic lipid membranes with one of the most typical classes of nanomaterials, citrate-coated AuNP, we addressed the cooperative nature of their interaction at the interface, which leads to AuNP clustering. The ensemble of optical (UV–Vis absorbance), structural (small-angle neutron and X-ray scattering) and surface (X-ray reflectivity, quartz crystal microbalance, atomic force microscopy) results, is consistent with a mechanistic hypothesis, where the citrate-lipid ligand exchange at the interface is the molecular origin of a multiscale cooperative behavior, which ultimately leads to the formation of clusters of AuNP on the bilayer. This mechanism, fully consistent with the data reported so far in the literature for synthetic bilayers, would shed new light on the interaction of engineered nanomaterials with biological membranes. The cooperative nature of ligand exchange at the AuNP-liposome interface, pivotal in determining clustering of AuNP, will have relevant implications for NP use in Nanomedicine, since NP will be internalized in cells as clusters, rather than as primary NP, with dramatic effects on their bioactivity.

Cysteamine-coated gold nanoparticles for bimodal colorimetric detection with inverse sensitivity: a proof-of-concept with lysozyme.

Cysteamine-coated gold nanoparticles (cysAuNPs) are positively charged as-synthesised and hence can interact with negatively charged DNA with ease. We have investigated the dependency of the particles' dispersion stage on different concentrations of lysozyme-binding aptamer (LBA). On top of the commonly reported phenomenon where cysAuNPs aggregate as the concentration of LBA increases, we observed that cysAuNPs redispersed after the amount of LBA achieved a certain threshold, dubbed as the critical redispersion concentration (CRC). By harnessing the aggregation and dispersion behaviour of cysAuNPs at LBA below and above the CRC, respectively, we have demonstrated a bimodal colorimetric aptasensor to detect lysozyme as a proof-of-concept study. Apart from being able to quantify the lysozyme in different ranges of concentrations with a visual change in colour, this aptasensor also demonstrated a novel concept of inverse sensitivity (i.e. higher signal with less analyte), leading to a 24-fold higher of signal-to-noise ratio (SNR), in comparison to the conventional sensors. The aptasensor can also selectively distinguish lysozyme and eliminate false results from other control proteins via both modes. The generalisability, as well as potential of cysAuNPs for bimodal colorimetric detection and inverse sensitivity behaviour have made this material an interesting alternative to citrate-coated AuNPs.

Synthesis, characterisation, and application of chamomile gold nanoparticles in molecular diagnostics: a new component for PCR kits

Various strategies have been suggested for successful amplification of the hard-to-amplify nucleic acids such as the inclusion of various chemical or biological materials in the in vitro nucleic acid amplification reactions, particularly polymerase chain reaction (PCR), and adjustment of the cycling programs. Although much efforts have been madefor improvements in the enhancement of polymerase chain reactions of high GC content nucleic acids, still significant challenges remain. In this study, the effects of citrate-coated AuNP and chamomile extract-coated AuNP (p-AuNP) on amplification of three genes with significant different GC percentage were evaluated. Owing to the enormous potential of gold nanoparticles in the enhancement of the PCR reactions, we showed the promising and consistent findings on the application of very dilute biocompatible chamomile-gold nanoparticles as safe and low-cost nanomaterials for molecular amplification of GC-rich DNA samples. We hypothesized that green AuNPs, which have different surface chemistry from cit-AuNPs, not only do not interfere with the PCR reactants but also are capable of enhancing PCR reactions. These results, for the first time, confirm the potential of using the green gold nanoparticles in the heat-assisted enzymatic in vitro reactions, suggesting Chamomile gold nanoparticles as reliable component of any PCR kits.

Development of theranostic active-targeting boron-containing gold nanoparticles for boron neutron capture therapy (BNCT)

Successful boron neutron capture therapy (BNCT) requires sufficient and specific delivery of boron atoms to malignant cells. Gold nanoparticles (AuNPs) have been used as a useful delivery system for selectively releasing cytotoxic payloads in the tumor. However, studies demonstrating the in vivo distribution or pharmacokinetics of boron-containing AuNPs via noninvasive imaging are lacking. This study aims to develop theranostic AuNP-boron cage assemblies (B-AuNPs) and evaluate its feasibility for BNCT. The commercial citrate-coated AuNPs were subjected to PEGylation, azide addition, and carborane modification on the surface. To further arm the AuNPs, we conjugated anti-HER2 antibody (61 IgG) with boron-containing PEGylated AuNPs to form 61-B-AuNPs. The diameter and radiolabeling efficiency of boron-containing AuNPs were determined by dynamic light scattering (DLS) and radio thin-layer chromatography (radio TLC), respectively. Noninvasive single-photon emission computed tomography (SPECT)/computed tomography (CT) imaging was performed to determine the pharmacokinetics of radioiodinated AuNPs in N87 gastric cancer xenografts, and the content of boron in tumor and muscle was assessed by inductively coupled plasma mass spectrometry (ICP-MS). After the 3-step modification, the diameter of B-AuNPs increased by ˜25 nm, and antibody conjugation did not affect the diameter of AuNPs. Radioactive iodine (I-123) was introduced in AuNPs by Click chemistry under copper catalysis. The radiolabeling efficiency of 123I-B-AuNPs and 123I-61-B-AuNPs was approximately 60 ± 5%. After purification, the radiochemical purity (RCP) of these NPs was greater than 90%. MicroSPECT/CT imaging showed that the tumor-to-muscle (T/M) ratio of 123I-B-AuNP-injected mice reached 1.91 ± 0.17 at 12 h post-injection, while that of 123I-61-B-AuNP-injected mice was 12.02 ± 0.94. However, the increased uptake of AuNPs by the thyroid was observed at 36 h after the administration of 123I-61-B-AuNPs, indicating antibody-mediated phagocytosis. The T/M ratio, assessed by ICP-MS, of B-AuNP- and 61-B-AuNP-injected mice was 4.91 ± 2.75 and 41.05 ± 11.15, respectively. We successfully developed detectable HER2-targeting boron-containing AuNPs with high RCP and an acceptable yield. Noninvasive imaging could be a valuable tool for the noninvasive determination of the pharmacokinetics of AuNPs and measurement of boron concentration in the tumor.

Probing the Aggregation and Immune Response of Human Islet Amyloid Polypeptides with Ligand-Stabilized Gold Nanoparticles.

The use of nanomaterials has recently become an emerging strategy against protein amyloidosis associated with a range of metabolic and brain diseases. To facilitate research in this area, here we first demonstrated the use of hyperspectral imaging (HSI) and COMSOL simulations for reporting the aggregation of human islet amyloid polypeptides (IAPPs), a hallmark of type 2 diabetes, as well as the physical interactions between the peptide and gold nanoparticles (AuNPs) grafted with citrate and poly(ethylene glycol) (PEG400 and PEG3000). We found a distinct anticorrelation between increased IAPP aggregation and decreased spectral red shifts incurred in the AuNP plasmonic resonance. Moreover, Jurkat cells exposed to IAPP and AuNPs were characterized by quantifying their cytokine secretions with a localized surface plasmon resonance (LSPR) immunoassay, where a peak response was registered for the most toxic IAPP oligomers and most suppressed by citrate-coated AuNPs. This study demonstrated the potential of using HSI and LSPR as two new platforms for the facile examination of protein aggregation and their induced immune response associated with amyloid diseases.

Functional, Degradable Zwitterionic Polyphosphoesters as Biocompatible Coating Materials for Metal Nanostructures.

A zwitterionic polyphosphoester (zPPE), specifically l-cysteine-functionalized poly(but-3-yn-1-yloxy)-2-oxo-1,3,2-dioxaphospholane (zPBYP), has been developed as a poly(ethylene glycol) (PEG) alternative coating material for gold nanoparticles (AuNPs), the most extensively investigated metal nanoparticulate platform toward molecular imaging, photothermal therapy, and drug delivery applications. Thiol-yne conjugation of cysteine transformed an initial azido-terminated and alkynyl-functionalized PBYP homopolymer into zPBYP, offering hydrolytic degradability, biocompatibility, and versatile reactive moieties for installation of a range of functional groups. Despite minor degradation during purification, zPPEs were able to stabilize AuNPs presumably through multivalent interactions between combinations of the side chain zwitterions (thioether and phosphoester groups of the zPPEs with the AuNPs). 31P NMR studies in D2O revealed ca. 20% hydrolysis of the phosphoester moieties of the repeat units had occurred during the workup and purification by aqueous dialysis at pH 3 over ca. 1 d, as observed by the 31P signal of the phosphotriesters resonating at ca. -0.5 to -1.7 shifting downfield to ca. 1.1 to -0.4 ppm, attributed to transformation to phosphates. Further hydrolysis of side chain and backbone units proceeded to an extent of ca. 75% over the next 2 d in nanopure water (pH 5-6). The NMR degradation results were consistent with the broadening and red-shift of the surface plasmon resonance (SPR) observed by UV-vis spectroscopy of the zPPE-coated AuNPs in water over time. All AuNP formulations in this study, including those with citrate, PEG, and zPPE coatings, exhibited negligible immunotoxicity, as determined by cytokine overexpression in the presence of the nanostructures relative to those in cell culture medium. Notably, the zPPE-coated AuNPs displayed superior antifouling properties, as assessed by the extent of cytokine adsorption relative to both the PEGylated and citrate-coated AuNPs. Taken together, the physicochemical and biological evaluations of zPPE-coated AuNPs in conjunction with PEGylated and citrate-coated analogues indicate the promise of zPPEs as favorable alternatives to PEG coatings, with negligible immunotoxicity, good antifouling performance, and versatile reactive groups that enable the preparation of highly tailored nanomaterials for diverse applications.

Novel Magnetic Microprobe with Benzoboroxole-Modified Flexible Multisite Arm for High-Efficiency cis-Diol Biomolecule Detection.

With regard to regulating a variety of biological events, including molecular recognition, signal transduction, cell adhesion, and immune response, cis-diol biomolecules, such as saccharides and glycoproteins, play vital roles. However, saccharides and glycoproteins in living systems usually exist in very low abundance, along with abundant interfering components. High-efficiency detection of saccharides and glycoproteins is a challenging yet highly impactful area of research. Herein, we reported a novel magnetic microprobe with a benzoboroxole-modified flexible multisite arm (PEG 2000-grafted PAMAM dendrimers; the microprobe was denoted as BFMA-MNP) for high-efficiency saccharides detection. The extraction capacity was significantly improved by ∼2 orders of magnitude, because of the integration of the enhanced hydrophilicity and multivalency effects in benzoboroxoles and the enhanced accessibility of the binding sites within the PEG 2000-grafted PAMAM dendrimers. As a result, the proposed approach possessed several advantages, compared with previous boronic acid-based methods, including ultrahigh sensitivity (limit of detection was <1 ng/mL), wide linear range (ranged from 0.5 μM to 2000 μM), and applicable in physiological pH condition. Furthermore, we established a general BFMA-MNP/glycoproteins/AuNPs sandwich assay to realize the visual glycoprotein qualitative screening for the first time. The unique sandwich assay possessed the dual nature of the magnetic separation by BFMA-MNPs and specific coloration by citrate-coated AuNPs. This visual sandwich assay enabled fast differentiation of the existence of glycoproteins in complicated samples without any advanced instruments. We believe the proposed BFMA-MNP microprobe herein will advance the ideas to detect and identify trace saccharides and glycoproteins in important fields such as glycomics and glycoproteomics.

Measurement of engineered nanoparticles in consumer products by surface-enhanced Raman spectroscopy and neutron activation analysis

There has been an increasing number of consumer and food products sold on the market that contain various engineered nanomaterials (ENMs) such as silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs). These nanomaterials possess novel physical and chemical properties that can be used for wide applications in agriculture and food safety. However, current analytical methods to detect and measure ENMs are time-consuming, labor-intensive, and expensive. The objective of this study was to develop a novel, simple, rapid, and accurate method to detect AgNPs and AuNPs in consumer products using surface-enhanced Raman spectroscopy (SERS). SERS measurement was conducted to detect AgNPs and AuNPs using an effective Raman indicator, 4-aminothiophenol (pATP). The pATP can strongly bind onto nanoparticles, generating greatly enhanced Raman signals that can be used for measurement. The pATP was combined with Ag or Au stock solution, AgNO3, citrate-coated AgNPs, citrate-coated AuNPs, AuCl, AgNPs, AuNPs, and five commercial products to study the differences in their SERS spectral data. The observed spectra of AgNPs and AuNPs have similar peaks at ~ 390, ~ 1087, and ~ 1590 cm−1 that can be attributed to the C–S stretching vibration, C–C stretching vibration, and C–H stretching vibration, respectively. Neutron activation analysis (NAA) and electron microscopy was used to characterize and quantify AgNPs and AuNPs in the consumer products. The results demonstrate that SERS method in combination with NAA can be an effective method for detection of ENMs, and it can easily distinguish AgNPs and AuNPs from other non-nanoparticle species in the complex matrices.

Understanding Protein Structure Deformation on the Surface of Gold Nanoparticles of Varying Size.

Gold nanoparticles (AuNPs) have been of recent interest due to their unique optical properties and their biocompatibility. Biomolecules spontaneously adsorb to their surface, a trait that could potentially be exploited for drug targeting. Currently, it is unclear whether protein-AuNP interactions at the nanoparticle surface are dependent on nanoparticle size. In this work, we investigate whether varying surface curvature can induce protein unfolding and multilayer binding in citrate-coated AuNPs of various sizes. A recently developed NMR-based approach was utilized to determine the adsorption capacity, and protein NMR spectra were compared to determine whether nanoparticle size influences protein interactions at the surface. In addition, transmission electron microscopy (TEM) and dynamic light scattering (DLS) were employed to corroborate the NMR studies. Over a broad range of AuNP sizes (14-86 nm), we show that adsorption capacity can be predicted by assuming that proteins are compact and globular on the nanoparticle surface. Additionally, roughly one layer of proteins is adsorbed regardless of AuNP size. Our results hold for two proteins of significantly different sizes, GB3 (6 kDa) and bovine carbonic anhydrase (BCA, 29 kDa). However, the unstable drkN SH3 domain (ΔḠ0 ≈ 0, 7 kDa) does not appear to follow the same trend seen for stable, globular proteins. This observation suggests that unstable proteins can deform significantly when bound to AuNP surfaces. Taken together, the results of this work can be used to improve our knowledge of the mechanism of protein-AuNP interactions to optimize their use in the biomedical field.

Electrostatic Interactions and Protein Competition Reveal a Dynamic Surface in Gold Nanoparticle-Protein Adsorption.

Gold nanoparticle- (AuNP-) protein conjugates are potentially useful in a broad array of diagnostic and therapeutic applications, but the physical basis of the simultaneous adsorption of multiple proteins onto AuNP surfaces remains poorly understood. Here, we investigate the contribution of electrostatic interactions to protein-AuNP binding by studying the pH-dependent binding behavior of two proteins, GB3 and ubiquitin. For both proteins, binding to 15-nm citrate-coated AuNPs closely tracks with the predicted net charge using standard pKa values, and a dramatic reduction in binding is observed when lysine residues are chemically methylated. This suggests that clusters of basic residues are involved in binding, and using this hypothesis, we model the pKa shifts induced by AuNP binding. Then, we employ a novel NMR-based approach to monitor the binding competition between GB3 and ubiquitin in situ at different pH values. In light of our model, the NMR measurements reveal that the net charge, binding association constant, and size of each protein play distinct roles at different stages of protein adsorption. When citrate-coated AuNPs and proteins first interact, net charge appears to dominate. However, as citrate molecules are displaced by protein, the surface chemistry changes, and the energetics of binding becomes far more complex. In this case, we observed that GB3 is able to displace ubiquitin at intermediate time scales, even though it has a lower net charge. The thermodynamic model for binding developed here could be the first step toward predicting the binding behavior in biological fluids, such as blood plasma.

Zeta-potential data reliability of gold nanoparticle biomolecular conjugates and its application in sensitive quantification of surface absorbed protein

Zeta potentials (ZP) of gold nanoparticle bioconjugates (AuNP-bios) provide important information on surface charge that is critical for many applications including drug delivery, biosensing, and cell imaging. The ZP measurements (ZPMs) are conducted under an alternative electrical field at a high frequency under laser irradiation, which may strongly affect the status of surface coating of AuNP-bios and generate unreliable data. In this study, we systemically evaluated the ZP data reliability (ZPDR) of citrate-, thiolated single stranded DNA-, and protein-coated AuNPs mainly according to the consistence of ZPs in the repeated ZPMs and the changes of the hydrodynamic size before and after the ZPMs. We found that the ZPDR was highly dependent on both buffer conditions and surface modifications. Overall, the higher ionic strength of the buffer and the lower affinity of surface bounders were related with the worse ZPDR. The ZPDR of citrate-coated AuNP was good in water, but bad in 10mM phosphate buffer (PB), showing substantially decrease of the absolute ZP values after each measurement, probably due to the electrical field facilitated adsorption of negatively charged phosphate ions on AuNPs. The significant desorption of DNAs from AuNP was observed in the PB containing medium concentration of NaCl, but not in PB. The excellent ZPDR of bovine serum albumin (BSA)-coated AuNP was observed at high salt concentrations and low surface coverage, enabling ZPM as an ultra-sensitive tool for protein quantification on the surface of AuNPs with a single molecule resolution.