The synthesis of a luminescent quantum cluster (QC) of gold with a quantum yield of ∼4% is reported. It was synthesized in gram quantities by the core etching of mercaptosuccinic acid protected gold nanoparticles by bovine serum albumin (BSA), abbreviated as AuQC@BSA. The cluster was characterized and a core of Au38 was assigned tentatively from mass spectrometric analysis. Luminescence of the QC is exploited as a "turn-off" sensor for Cu 2+ ions and a "turn-on" sensor for glutathione detection. Metal-enhanced luminescence (MEL) of this QC in the presence of silver nanoparticles is demonstrated and a ninefold maximum enhancement is seen. This is the first report of the observation of MEL from QCs. Folic acid conjugated AuQC@BSA was found to be internalized to a significant extent by oral carcinoma KB cells through folic acid mediated endocytosis. The inherent luminescence of the internalized AuQC@BSA was used in cell imaging. © 2010 Wiley-VCH Verlag GmbH &amp; Co. KGaA, Weinheim.

Pradeep Thalappil

Department Of Chemistry

BIOLABELING

Bovine serum albumins

CORE ETCHING

Enhanced luminescence

FOLIC ACIDS

Glutathiones

Gold Nanoparticles

GRAM QUANTITIES

IN-CELL

Mass spectrometric analysis

MERCAPTOSUCCINIC ACIDS

Metal ion sensing

QUANTUM CLUSTERS

Silver nanoparticles

Body fluids

etching

Gold compounds

Luminescence

Metal ions

Nanoparticles

Organic acids

Organic polymers

quantum yield

Sensors

Silver

Gold

Bovine serum albumin

cation

metal nanoparticle

Animal

article

Cattle

chemistry

Human

Animals

Cations

Humans

Metal nanoparticles

Serum Albumin, Bovine

IIT Madras is a public technical and research university located in Chennai, Tamil Nadu. Founded in 1959, it is recognised as an Institute of National Importance.

IIT Madras has been ranked as the top engineering institute in India for four years in a row by the National Institutional Ranking Framework of the MHRD

It currently offers undergraduate, postgraduate and research degrees across 16 disciplines in Engineering, Sciences, Humanities and Management. About 596 faculty belonging to science and engineering departments and centres of the Institute are engaged in teaching, research and industrial consultancy.

IIT Madras

Chemistry - A European Journal

A new synthetic protocol is introduced which preserves the secondary structure of protecting proteins encapsulating a luminescent atomically-precise silver cluster. This was achieved by using a preformed triphenylphosphine (TPP)-protected silver cluster as the precursor forming bovine serum albumin (BSA)- and human serum albumin (HSA)-protected Ag18 clusters. This is the first example of the formation of luminescent protein-protected clusters in a neutral medium, without using any reducing agent, which results in minimal alteration of the protein structure during cluster growth. The cluster formed showed exceptional stability, unlike other silver clusters of this class. The formation of these red luminescent clusters was visualized by UV-vis and photoluminescence spectroscopy. The identification of Ag18 core was made through matrix-assisted laser desorption ionization mass spectrometry (MALDI MS), and a plausible mechanism of the formation was identified by monitoring the systematic growth of the cluster core by time-dependent MALDI MS experiments and electrospray ionization mass spectrometry (ESI MS) of the reaction mixture. The cluster was successfully employed as a luminescent probe for cancer cell imaging. Retention of protein conformation in the clusters was confirmed through circular dichroism (CD) spectroscopy, and the same was reflected in the retention of 89% of the esterase activity of BSA in the Ag18@BSA clusters synthesized by this method, compared to only 28.7% for AgQC@BSA clusters synthesized using previous protocols, conducted in basic medium. © 2019 American Chemical Society.

Journal of Physical Chemistry C

Internalization of a Preformed Atomically Precise Silver Cluster in Proteins by Multistep Events and Emergence of Luminescent Counterparts Retaining Bioactivity

Hydrogen/deuterium exchange mass spectrometry was employed to probe the conformational changes in lysozyme (Lyz) during the course of formation of a protein-protected atomically precise Au8 cluster. MALDI MS showed the protein, Lyz, to be present in a denatured state in the cluster. Detailed ESI MS analysis of the Au-attached Lyz adducts, an intermediate of cluster formation, confirmed that these conformational changes are brought about by Au-S bond formation and a similar conformation is retained in the final cluster. These results were supported by computational results, which showed an increase in solvent accessible surface area upon the formation of the adducts. Infrared spectroscopy established that change in the rate as well as the extent of the hydrogen/deuterium exchange observed in the cluster was due to the change in the amide II region of the encapsulating protein. Hydrogen/deuterium exchange ESI MS of Cu adducts of Lyz showed a lower degree of denaturation than their Au counterparts. XPS analysis revealed that Cu binds differently to Lyz than it does with Au, which is likely because of the stronger soft-soft Au-S interaction. Alkali metal ion binding, on the other hand, does not affect the protein conformation because such ions do not affect the disulfide bonds. © 2019 American Chemical Society.

Conformational Changes of Protein upon Encapsulation of Noble Metal Clusters: An Investigation by Hydrogen/Deuterium Exchange Mass Spectrometry

We probed the interaction between Au38@BSA and various heavy metal ions using luminescence spectroscopy. Interestingly, Au38@BSA showed luminescence enhancement upon interaction with Cd2+ and Pb2+ at concentrations higher than 1 ppm, due to the formation of cluster aggregates. Such aggregates were detected by dynamic light scattering (DLS) and high resolution electron microscopy (HRTEM) studies. Luminescence enhancement of Au38@BSA in the presence of Cd2+ was due to the interaction of Cd2+ with the cluster core, while Pb2+-induced luminescence enhancement was due to BSA-Pb2+ interaction. Observations were further supported by X-ray photoelectron spectroscopy (XPS) studies. This kind of phenomenon has been observed in protein protected clusters for the first time. We believe that such metal-ion-induced luminescence enhancement can be used to synthesize cluster systems with enhanced optical properties and different ion-cluster interactions can be used to develop metal ion sensors using Au38@BSA. © 2019 American Chemical Society.

Metal-Ion-Induced Luminescence Enhancement in Protein Protected Gold Clusters

Ion mobility mass spectrometry studies on Aun-Lyz adducts showed gradual unfolding of the protein structure during binding of Au+ to the protein. The change of the charge state envelope in Aun-Lyz from that of Lyz in ESI MS data confirmed the relaxation of the protein structure. This Au+ binding occurs at cysteine sites through the breakage of disulfide bonds and this ruptures the H-bonded folded network structure of the protein leading to ∼30% change in helicity. Nearly 15% loss in the total H-bonding occurred during the attachment of 8 Au to the protein as calculated by a molecular dynamics simulation. Different Aun-Lyz structures were simulated, which confirmed significant unfolding of the protein. The structural insights were used to understand similar unfolding in the solution state as seen via circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy. This open structure is indeed necessary to accommodate a cluster core inside a protein cavity during luminescent cluster synthesis. These studies unambiguously establish noble metal binding-induced conformational changes of protein structures to accommodate the clusters. © 2017 American Chemical Society.

Gold-Induced Unfolding of Lysozyme: Toward the Formation of Luminescent Clusters

Atomically precise gold and silver clusters are a new class of sensitizers which can be used as substitutes for dyes in the classical dye-sensitized solar cells (DSCs). Here noble metal clusters protected by proteins and thiols (Au30@BSA, Au25SBB18, and Ag44MBA30) have been used for photovoltaic studies. These metal clusters were used as sensitizers for the photoanodes fabricated using TiO2 nanotubes (NTs) and the commercial P25 TiO2 nanoparticles. The TiO2, clusters and the solar cells were characterized by spectroscopy, microscopy, current-voltage (I-V) and incident photon-to-current conversion efficiency (IPCE) measurements. A systematic I-V study revealed a conversion efficiency of 0.35 % for the Au30@BSA sensitized solar cell made from TiO2 NTs which showed an IPCE maximum of 3 % at ∼ 400 nm. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

ChemistrySelect

Atomically Precise Noble Metal Clusters Harvest Visible Light to Produce Energy

Molecular-receptor-targeted imaging of folate receptor positive oral carcinoma cells using folic-acid-conjugated fluorescent Au25 nanoclusters(Au NCs) is reported. Highly fluorescent Au25 clusters were synthesized by controlled reduction of Au+ ions, stabilized in bovine serum albumin(BSA), using a green-chemical reducing agent, ascorbic acid (vitamin-C). For targeted-imaging-based detection of cancer cells, the clusters were conjugated with folic acid(FA) through amide linkage with the BSA shell. The bioconjugated clusters show excellent stability over a wide range of pH from 4 to 14 and fluorescence efficiency of ∼5.7% at pH 7.4 in phosphate buffer saline(PBS), indicating effective protection of nanoclusters by serum albumin during the bioconjugation reaction and cell-cluster interaction. The nanoclusters were characterized for their physico-chemical properties, toxicity and cancer targeting efficacy in vitro. X-ray photoelectron spectroscopy(XPS) suggests binding energies correlating to metal Au4f7/2∼83.97eV and Au4f5/2∼87.768eV. Transmission electron microscopy and atomic force microscopy revealed the formation of individual nanoclusters of size ∼1nm and protein cluster aggregates of size ∼8nm. Photoluminescence studies show bright fluorescence with peak maximum at ∼674nm with the spectral profile covering the near-infrared(NIR) region, making it possible to image clusters at the 700-800nm emission window where the tissue absorption of light is minimum. The cell viability and reactive oxygen toxicity studies indicate the non-toxic nature of the Au clusters up to relatively higher concentrations of 500νgml-1. Receptor-targeted cancer detection using Au clusters is demonstrated on FR+ve oral squamous cell carcinoma (KB) and breast adenocarcinoma cell MCF-7, where the FA-conjugated Au25 clusters were found internalized in significantly higher concentrations compared to the negative control cell lines. This study demonstrates the potential of using non-toxic fluorescent Au nanoclusters for the targeted imaging of cancer. © IOP Publishing Ltd.

Fulltext

Nanotechnology

Molecular-receptor-specific, non-toxic, near-infrared-emitting Au cluster-protein nanoconjugates for targeted cancer imaging

We synthesized fluorescent, porphyrin-anchored, Au 22 clusters in a single step, starting from well-characterized Au 25 clusters protected with glutathione (-SG) by a combined core reduction/ligand exchange protocol, at a liquid-liquid interface. The prepared cluster was characterized by UV/vis, photoluminescence, X-ray photoelectron spectroscopy (XPS), scanning electron microscopy, elemental analysis, and matrix-assisted laser desorption ionization mass spectrometry. The absence of a 672 nm intraband transition of Au 25 and the simultaneous emergence of new characteristic peaks at 520 and 635 nm indicate the formation of the Au 22 core. An increase in the binding energy of 0.4 eV in Au 4f core-level peaks confirmed the presence of a reduced core size. Quantitative XPS confirmed the Au/S ratio. The presence of a free base, tetraphenylporphyrin (H 2TPPOAS-), on the Au 22 core was confirmed by fluorimetric titrations with Cu 2+ and Zn 2+ ions. From all of these, the composition of the cluster was determined to be Au 22[(-SG) 15(-SAOPPTH 2) 2], which was supported by mass spectrometry and elemental analysis. We utilized the fluorescence nature of these water-soluble clusters for the fabrication of fluorescent patterns by soft lithography. The patterns were studied using tapping-mode atomic force microscopy and confocal fluorescence imaging. © 2009 American Chemical Society.

ACS Applied Materials and Interfaces

Functionalized Au 22 clusters: Synthesis, characterization, and patterning

Glutathione protected Au25 quantum clusters, exhibiting characteristic fluorescence, have been uniformly coated inside and outside of β-Ala-L-Ile dipeptide nanotubes. These coated structures have been imaged using the inherent fluorescence of Au25. Upon exposure to an electron beam, in a transmission electron microscope, the quantum clusters gradually transform to gold nanoparticles, of the metallic size regime. The nanoparticles grow to a size of 4.5 nm and thereafter the particle size is unaffected by electron beam exposure. The nanotubes are intact and this template is shown to control the uniformity of the size of the nanoparticles grown. The quantum clusters can be loaded selectively inside the tubes using capillarity of the nanotubes. The sizes of the nanoparticles grown are tuned using electron beam exposure. © 2009 The Royal Society of Chemistry.

Journal of Materials Chemistry

Size tuning of Au nanoparticles formed by electron beam irradiation of Au25 quantum clusters anchored within and outside of dipeptide nanotubes

A novel interfacial route has been developed for the synthesis of a bright-red-emitting new subnanocluster, Au23, by the core etching of a widely explored and more stable cluster, Au25SG18 (in which SG is glutathione thiolate). A slight modification of this procedure results in the formation of two other known subnanoclusters, Au22 and Au33. Whereas Au22 and Au23 are water soluble and brightly fluorescent with quantum yields of 2.5 and 1.3%, respectively, Au33 is organic soluble and less fluorescent, with a quantum yield of 0.1%. Au23 exhibits quenching of fluorescence selectively in the presence of Cu2+ ions and it can therefore be used as a metal-ion sensor. Aqueous- to organic-phase transfer of Au23 has been carried out with fluorescence enhancement. Sol-vent dependency on the fluorescence of Au23 before and after phase transfer has been studied extensively and the quantum yield of the cluster varies with the solvent used. The temperature response of Au23 emission has been demonstrated. The inherent fluorescence of Au23 was used for imaging human hepatoma cells by employing the avidin-biotin interaction. © 2009 Wiley-VCH Verlag GmbH &amp; Co. KGaA.

Bright, NIR-emitting Au23 from Au25: Characterization and applications including biolabeling

Ligand exchange offers an effective way to modify the properties of the recently prepared quantum clusters of gold. To tune optical and photoluminescence properties of one of the most stable quantum clusters of gold, AU25SG18 (SG-glutathione thiolate), we functionalized it by the exchange of - SG with functionalized -SG and with an altogether different ligand, namely, 3-mercapto-2-butanol (MB). The products were characterized by various techniques such as optical absorption (UV - vis), Fourier-transform infrared (FT - IR), nuclear magnetic resonance (NMR), X-ray photoelectron (XPS), and luminescence spectroscopies, mass spectrometry, and thermogravimetry (TG). Analyses of the TG data helped to establish the molecular composition of the products. Ligand exchange reaction was monitored by NMR spectroscopy, and it was found that the exchange reaction follows a first order kinetics. The XPS study showed that after the exchange reaction there was no change in the chemical nature of the metal core and binding energy values of Au 4f7/2 and 4f5/2, which are similar in both the parent and the exchanged products. Photoluminescence studies of these clusters, done in the aerated conditions, showed that the excitation spectrum of the MB-exchanged product is entirely different from the acetyl- and formyl-glutathione exchanged products. The inherent fluorescence and solid-state emission of these clusters were observed. This intense emission allows optical imaging of the material in the solid state. The emission is strongly temperature dependent. The synthesis of a diverse variety of clusters and their chemical stability and intense luminescence offer numerous applications in areas such as energy transfer, sensors, biolabeling, and drug delivery. © 2008 American Chemical Society.

Ligand exchange of Au25SG18 leading to functionalized gold clusters: Spectroscopy, kinetics, and luminescence

Luminescent quantum clusters of gold in bulk by albumin-induced core etching of nanoparticles: Metal ion sensing, metal-enhanced luminescence, and biolabeling

Journal	Chemistry - A European Journal
ISSN	09476539
Open Access	No