Fluorescent silver nanoclusters as antibody label in a competitive immunoassay for the complement factor H

Valencia, Eva; Cruz-Alonso, María; Álvarez, Lydia; González‐Iglesias, Héctor; Fernández, Beatriz; Pereiro, Rosario

doi:10.1007/s00604-019-3554-y

Cited by 14 publications

(12 citation statements)

References 40 publications

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“…Next, MNCs were characterised by fluorescence, HR-TEM and ICP-MS measurements. HR-TEM images showed in the three cases MNCs with a spherical metal nucleus and FCC crystal structure [26,29,30]. As described elsewhere [26,29,30], the average particle size determined for each type of MNCs was: 1.5 ± 0.6 nm for PtNCs, 2.20 ± 0.04 nm for AuNCs, and 2.9 ± 0.1 nm for AgNCs.…”

Section: Synthesis Of Metal Nanoclusterssupporting

confidence: 56%

“…To employ PtNCs (and MNCs in general) as labels for examining protein distribution in biological tissues, an additional step is necessary after the binding of the Ab to the MNCs in the preparation of the labelled immunoprobes to minimise non-specific interactions with tissues. Primary amines, such as hydroxylamine have been successfully proved to block carboxylic groups of AuNCs and AgNCs [26,27]. In the present work three molar ratios of PtNCs:hydroxylamine (1:100, 1:50 and 1:20) were evaluated for reducing nonspecific PtNCs:anti-h-APP binding in the eye sections (Fig.…”

Section: Optimisation Of the Bioconjugation And Ihc Procedures Using Ptncs As Labelsmentioning

confidence: 99%

“…The syntheses of the three thiolated MNCs (PtNCs, AuNCs and AgNCs) were carried out following the protocols described elsewhere [26,29,30]. A similar bottom-up procedure was used in all cases: the metal precursor (salt of the metal with a high oxidation state) is dissolved in basic aqueous media (adjusted with a NaOH solution) and subsequently reduced by NaBH4 under the presence of the stabilising agent (lipoic acid).…”

Section: Synthesis Of Metal Nanoclustersmentioning

confidence: 99%

“…These nanoparticles contain thousands of metal atoms, but serious non-specific adsorptions have been described. The use of metal nanoclusters (MNCs) with diameters below 3 nm, typically containing a few hundreds of metal atoms, offers lower risk of blocking recognition sites of the biomolecules [24], more efficient penetration within the biological tissue during the IHC procedure, as well as high amplification [25][26][27]. AuNCs and AgNCs have been successfully investigated for specific proteins bioimaging by LA-ICP-MS [25][26][27].…”

Section: Introductionmentioning

confidence: 99%

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Multiplex bioimaging of proteins-related to neurodegenerative diseases in eye sections by laser ablation - Inductively coupled plasma – Mass spectrometry using metal nanoclusters as labels

Lores-Padín

Fernández

Álvarez

et al. 2021

Talanta

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Simultaneous determination of proteins with micrometric resolution is a significant challenge. In this study, laser ablation (LA) inductively coupled plasma -mass spectrometry (ICP-MS) was employed to quantify the distribution of proteins associated to the eye disease age-related macular degeneration (AMD) using antibodies labelled with three different metal nanoclusters (MNCs). PtNCs, AuNCs and AgNCs contain hundreds of metal atoms and were used to detect metallothionein 1/2 (MT1/2), complement factor H (CFH) and amyloid precursor protein (APP) in retina, ciliary body, retinal pigment epithelium (RPE), choroid and sclera from human cadaveric eye sections. First, the labelling of MNCs bioconjugated primary antibodies (Ab) was optimised following an immunolabelling protocol to avoid the non-specific interaction of MNCs with the tissue.Then, the LA and ICP-MS conditions were studied to obtain high-resolution images for the simultaneous detection of the three labels at the same tissue section. A significant signal amplification was found when using AuNCs, AgNCs and PtNCs labelled Ab of 310, 723 and 1194 respectively. After the characterisation of MNCs labelled immunoprobes, the antibody labelling was used for quantitative determination of MT1/2, CFH and APP in the RPE-choroid-sclera, where accumulation of extracellular deposits related to AMD was observed, suggesting that this method is fully suitable for the simultaneous detection and quantification of at least three different proteins.

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Section: Synthesis Of Metal Nanoclusterssupporting

confidence: 56%

Section: Optimisation Of the Bioconjugation And Ihc Procedures Using Ptncs As Labelsmentioning

confidence: 99%

Section: Synthesis Of Metal Nanoclustersmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Multiplex bioimaging of proteins-related to neurodegenerative diseases in eye sections by laser ablation - Inductively coupled plasma – Mass spectrometry using metal nanoclusters as labels

Lores-Padín

Fernández

Álvarez

et al. 2021

Talanta

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“…Gold is, by far, the most common metal employed for the synthesis of fluorescent NCs [13]. Silver NCs [14,15] and copper NCs [16] are also frequently investigated. Nowadays, the synthesis of fluorescent NCs of other metals is pursued, such as iridium [17] and molybdenum [18].…”

Section: Introductionmentioning

confidence: 99%

Bimodal determination of immunoglobulin E by fluorometry and ICP-MS by using platinum nanoclusters as a label in an immunoassay

et al. 2019

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The authors describe the use of platinum nanoclusters (PtNCs) as bimodal labels in a competitive immunoassay for immunoglobulin E (IgE). Both fluorometry and inductively coupled plasmamass spectrometry (ICP-MS) are used. Optimization of the PtNCs synthesis process using lipoic acid as ligand was carried out. The time for synthesis and the effect of NaOH added to the PtNCs precursor mixture was optimized with the aim to obtain PtNCs with strong fluorescence and low size dispersity. Maximal fluorescence was obtained at excitation/emission wavelengths of 455/620 nm. The average diameter (1.5 nm) and crystal structure (face-centered cubic structure) of the PtNCs were determined by HR-TEM. It was calculated that each PtNC contains 116 Pt atoms at average. Labelling of the antibody (Ab) against IgE with PtNCs was optimized in terms of recognition capabilities and fluorescence signal. A molar ratio (Ab:PtNCs) of 1:11 is found to be best. A competitive immunoassay for IgE was developed and detection was carried out by using both ICP-MS (by measuring 195 Pt) and fluorometry. The limit of detection (LOD) of the fluoroimmunoassay is 0.6 ng mL-1 of IgE. The LOD of the ICP-MS method is as low as 0.08 ng mL-1. The method was evaluated by analyzing four (spiked) serum samples by ICP-MS. No sample pretreatment excepting dilution is needed. Results compared favorably with those obtained by a commercial ELISA kit.

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Bioactive Luminescent Silver Clusters Confined in Zeolites Enable Quick and Wash‐Free Biosensing

García‐Guzmán,

Morales‐Narváez,

Coutino‐Gonzalez

2023

Angewandte Chemie

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The simultaneous capture and detection of biomolecules is crucial for revolutionizing bioanalytical platforms in terms of portability, response time and cost‐efficiency. Herein, we demonstrate how the sensitivity to external stimuli and changes in the local electronic environment of silver clusters lead to an advantageous biosensing platform based on the fluorometric response of bioactive luminescent silver clusters (BioLuSiC) confined in faujasite X zeolites functionalized with antibodies. The photoluminescence response of BioLuSiC was enhanced upon immunocomplex formation, empowering a wash‐free and quick biodetection system offering optimal results from 5 min. Proteins and pathogens (immunoglobulin G and Escherichia coli) were targeted to demonstrate the biosensing performance of BioLuSiC, and a human serum titration assay was also established. BioLuSiC will pave the way for innovative bioanalytical platforms, including real‐time monitoring systems, point‐of‐care devices and bioimaging techniques.

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Fluorescent silver nanoclusters as antibody label in a competitive immunoassay for the complement factor H

Cited by 14 publications

References 40 publications

Multiplex bioimaging of proteins-related to neurodegenerative diseases in eye sections by laser ablation - Inductively coupled plasma – Mass spectrometry using metal nanoclusters as labels

Multiplex bioimaging of proteins-related to neurodegenerative diseases in eye sections by laser ablation - Inductively coupled plasma – Mass spectrometry using metal nanoclusters as labels

Bimodal determination of immunoglobulin E by fluorometry and ICP-MS by using platinum nanoclusters as a label in an immunoassay

Bioactive Luminescent Silver Clusters Confined in Zeolites Enable Quick and Wash‐Free Biosensing

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