Functionalization of colloidal nanoparticles with a discrete number of ligands based on a “HALO-bioclick” reaction

Garbujo, Stefania; Galbiati, Elisabetta; Salvioni, Lucia; Mazzucchelli, Matteo; Frascotti, Gianni; Sun, Xing; Megahed, Saad; Feliu, Neus; Prosperi, Davide; Parak, Wolfgang J.; Colombo, Miriam

doi:10.1039/d0cc04355a

Cited by 9 publications

(10 citation statements)

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“…Poly-(isobutylene-alt-maleic anhydride)- graft -dodecyl polymer (PMA) was synthetized following a previously published protocol [ 35 ] with some modifications. In detail, 3.084 g (20 mmol expressed as in terms of monomer units) of poly(isobutylene-alt-maleic anhydride) (average molecular weight (Mw) ~6000 g/mol) was dissolved in 100 mL of anhydrous tetrahydrofuran (THF).…”

Section: Methodsmentioning

confidence: 99%

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The Role of Polymeric Coatings for a Safe-by-Design Development of Biomedical Gold Nanoparticles Assessed in Zebrafish Embryo

et al. 2021

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In the biomedical field, gold nanoparticles (GNPs) have attracted the attention of the scientific community thanks to their high potential in both diagnostic and therapeutic applications. The extensive use of GNPs led researchers to investigate their toxicity, identifying stability, size, shape, and surface charge as key properties determining their impact on biological systems, with possible strategies defined to reduce it according to a Safe-by-Design (SbD) approach. The purpose of the present work was to analyze the toxicity of GNPs of various sizes and with different coating polymers on the developing vertebrate model, zebrafish. In particular, increasing concentrations (from 0.001 to 1 nM) of 6 or 15 nm poly-(isobutylene-alt-maleic anhydride)-graft-dodecyl polymer (PMA)- or polyethylene glycol (PEG)-coated GNPs were tested on zebrafish embryos using the fish embryo test (FET). While GNP@PMA did not exert significant toxicity on zebrafish embryos, GNP@PEG induced a significant inhibition of embryo viability, a delay of hatching (with the smaller size NPs), and a higher incidence of malformations, in terms of tail morphology and eye development. Transmission electron microscope analysis evidenced that the more negatively charged GNP@PMA was sequestered by the positive charges of chorion proteins, with a consequent reduction in the amount of NPs able to reach the developing embryo and exert toxicological activity. The mild toxic response observed on embryos directly exposed to GNP@PMA suggest that these NPs are promising in terms of SbD development of gold-based biomedical nanodevices. On the other hand, the almost neutral GNP@PEG, which did not interact with the chorion surface and was free to cross chorion pores, significantly impacted the developing zebrafish. The present study raises concerns about the safety of PEGylated gold nanoparticles and contributes to the debated issue of the free use of this nanotool in medicine and nano-biotechnologies.

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Section: Methodsmentioning

confidence: 99%

“…First, 6 nm and 15 nm GNPs suspended in chloroform were transferred to an aqueous solution by wrapping the amphiphilic polymer PMA around their surface, as described in a previously published protocol [ 35 ]. An aliquot of polymer (Vp) at a concentration of Cp = 0.5 M was added to 6 nm and 15 nm GNPs suspended in chloroform.…”

Section: Methodsmentioning

confidence: 99%

The Role of Polymeric Coatings for a Safe-by-Design Development of Biomedical Gold Nanoparticles Assessed in Zebrafish Embryo

et al. 2021

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“…This technology overcomes the current limitations of traditional protein tagging platforms, as it can be applied to protein isolation and purification, studies of protein synthesis and degradation, analyses of protein function, studies of protein-protein and protein-DNA interactions, and molecular and cellular imaging (Encell et al, 2012;Merrill et al, 2019;Cattoglio et al, 2020;Freitas et al, 2021;Minner-Meinen et al, 2021). Furthermore, novel technologies have been developed for tumor diagnosis and treatment involving the linkage of dehalogenase fused with cancer cell recognition peptides to multifunctional nanoparticles (Garbujo et al, 2020).…”

Section: Other Fieldsmentioning

confidence: 99%

Mini Review: Advances in 2-Haloacid Dehalogenases

et al. 2021

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The 2-haloacid dehalogenases (EC 3.8.1.X) are industrially important enzymes that catalyze the cleavage of carbon–halogen bonds in 2-haloalkanoic acids, releasing halogen ions and producing corresponding 2-hydroxyl acids. These enzymes are of particular interest in environmental remediation and environmentally friendly synthesis of optically pure chiral compounds due to their ability to degrade a wide range of halogenated compounds with astonishing efficiency for enantiomer resolution. The 2-haloacid dehalogenases have been extensively studied with regard to their biochemical characterization, protein crystal structures, and catalytic mechanisms. This paper comprehensively reviews the source of isolation, classification, protein structures, reaction mechanisms, biochemical properties, and application of 2-haloacid dehalogenases; current trends and avenues for further development have also been included.

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“…Compared to conventional fluorophores, QDs exhibit enhanced photophysical properties that result in improved sensitivity for biosensing and bioimaging applications (13,(19)(20)(21). While initial concerns regarding toxicity and general biological stability have limited the broad adoption of QDs in the biomedical field, a robust body of work has emerged that shows excellent stability and specificity of biological probes incorporating QDs (22)(23)(24)(25)(26)(27)(28)(29).…”

Section: Introductionmentioning

confidence: 99%

A Quantum Dot Biomimetic for SARS-CoV-2 to Interrogate Dysregulation of the Neurovascular Unit Relevant to Brain Inflammation

Chiang

Stout

Yanchik-Slade

et al. 2022

Preprint

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Despite limited evidence for competent infection and viral replication of SARS-CoV-2 in the central nervous system (CNS), neurologic dysfunction is a common post-acute medical condition reported in “recovered” COVID-19 patients. To identify a potential noninfectious route for SARS-CoV-2-mediated neurological damage, we constructed colloidal nanocrystal quantum dots linked to micelles decorated with spike protein (COVID-QDs) as a biomimetic to interrogate how blood-brain barrier (BBB) dysregulation may subsequently induce neuroinflammation in the absence of infection. In transwell co-culture of endothelial bEnd.3 monolayers and primary neuroglia, we exposed only the bEnd.3 monolayers to COVID-QDs and examined by fluorescence microscopy whether such treatment led to (i) increased inflammation and leakage across the bEnd.3 monolayers, (ii) permeability of the COVID-QDs across the monolayers, and (iii) induction of neuroinflammation in neuroglial cultures. The results of our study provide evidence of neuroinflammatory hallmarks in cultured neurons and astrocytes without direct exposure to SARS-CoV-2-like nanoparticles. Additionally, we found that pre-treatment of our co-cultures with a small-molecule, broad-spectrum inhibitor of mixed lineage and leucine rich repeat kinases led to reversal of the observed dysregulation in endothelial monolayers and resulted in neuroglial protection. The results reported here may serve to guide future studies into the potential mechanisms by which SARS-CoV-2 mediates neurologic dysfunction.

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Functionalization of colloidal nanoparticles with a discrete number of ligands based on a “HALO-bioclick” reaction

Abstract: A recombinant HALO-GFP fusion protein was designed and isolated to demonstrate the feasibility of controlling the number and orientation of protein ligands to be conjugated on colloidal gold nanoparticles. AuNPs...

Cited by 9 publications

References 28 publications

The Role of Polymeric Coatings for a Safe-by-Design Development of Biomedical Gold Nanoparticles Assessed in Zebrafish Embryo

The Role of Polymeric Coatings for a Safe-by-Design Development of Biomedical Gold Nanoparticles Assessed in Zebrafish Embryo

Mini Review: Advances in 2-Haloacid Dehalogenases

A Quantum Dot Biomimetic for SARS-CoV-2 to Interrogate Dysregulation of the Neurovascular Unit Relevant to Brain Inflammation

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