Oligonucleotide solid-phase synthesis on fluorescent nanoparticles grafted on controlled pore glass

Crozals, Gabriel De; Farre, Carole; Hantier, Grégoire; Léonard, Didier; Marquette, Christophe A.; Mandon, Céline A.; Marmuse, Laurence; Louis, Cédric; Toulmé, Jean‐Jacques; Billotey, Claire; Janier, Marc; Chaix, Carole

doi:10.1039/c2ra22077f

Cited by 9 publications

(17 citation statements)

References 32 publications

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“…Si-NPs was functionalized by an innovative solid-phase synthesis technology which enables functionalization of nanosized particles with DNA fragments, as reported previously (Bonnet et al 2018;De Crozals et al 2012). Briefly, the nanoparticle immobilization onto controlled pore glass (CPG) allows a very high functionalization with a modified oligonucleotidic based linker.…”

Section: Si-np Synthesis Labeling and Quantificationmentioning

confidence: 99%

Highly Sensitive Detection ofCampylobacter spp.in Chicken Meat using a Silica Nanoparticle Enhanced Dot Blot DNA Biosensor

Vizzini

Manzano

Farre

et al. 2020

Preprint

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AbstractPaper-based DNA biosensors are powerful tools in point-of-care diagnostics since they are affordable, portable, user friendly, rapid and robust. However, their sensitivity is not always as high as required to enable DNA quantification. To improve the response of standard dot blots, we have applied a new enhancement strategy that increases the sensitivity of assays based on the use of biotinylated silica-nanoparticles (biotin-Si-NPs). After immobilization of a genomic Campylobacter DNA onto a paper membrane, and addition of a biotinylated-DNA detection probe, the hybridization was evidenced using streptavidin-conjugated to horseradish peroxidase (HRP) in the presence of luminol and H2O2. Replacement of the single biotin by the biotin-Si-NPs boosted in average a 30 fold the chemiluminescent read-out of the biosensor. The characterization of biotin-Si-NPs into a paper with immobilized DNA was done using a scanning electron microscope. A limit of detection of 3 pg/μL of DNA, similar to the available qPCR kits, is achieved but being cheaper, easier and avoiding the inhibition of DNA polymerase by molecules from the food matrices. We demonstrated that the new dot blot coupled to biotin-Si-NPs successfully detected Campylobacter from naturally contaminated chicken meat, without a need for PCR step. Hence, such enhanced dot blot paves the path to the development of a portable and multiplex paper based platform for point-of-care screening of chicken carcasses for Campylobacter.

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Section: Si-np Synthesis Labeling and Quantificationmentioning

confidence: 99%

Highly Sensitive Detection ofCampylobacter spp.in Chicken Meat using a Silica Nanoparticle Enhanced Dot Blot DNA Biosensor

Vizzini

Manzano

Farre

et al. 2020

Preprint

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“…Rhodamine-coated silica nanoparticles were functionalized by a supported synthesis strategy developed by our group [31]. The functionalization was achieved in a DNA/RNA synthesizer using phosphoramidite chemistry.…”

Section: Biotin Coating On Silica Npmentioning

confidence: 99%

“…Silica nanoparticles of 50 nm diameter were functionalized on surface with an oligonucleotidic strand bearing a biotin group at its extremity. The method used for NP functionalization was based on a solid-phase synthesis technology previously reported by our group [31]. Nanoparticles (NP) were first immobilized on a control pore glass (CPG) support.…”

Section: Biotin-np Developmentmentioning

confidence: 99%

“…Experiments were performed in the lysis buffer complemented with saliva to simulate clinical samples. Fifty-nanometre-diameter silica NP were functionalized via phosphoramidite chemistry, using a strategy of supported synthesis, adapting a protocol previously described by our group [31]. This well-controlled and reproducible method proved effective for NP functionalization with methylene bluemodified oligonucleotides [25] and proteins [32].…”

Section: Introductionmentioning

confidence: 99%

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Specific and sensitive detection of Influenza A virus using a biotin-coated nanoparticle enhanced immunomagnetic assay

et al. 2020

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This study reports the development of a sensitive magnetic bead-based enzyme-linked immunoassay (MELISA) for the panreactive detection of the Influenza A virus. The assay combines immunomagnetic beads and biotin-nanoparticle-based detection to quantify a highly conserved viral nucleoprotein in virus lysates. At the capture step, monoclonal antibody-coated magnetic microbeads were used to bind and concentrate the nucleoprotein in samples. The colorimetric detection signal was amplified using biotinylated silica nanoparticles (NP). These nanoparticles were functionalized on the surface with short DNA spacers bearing biotin groups by an automated supported synthesis method performed on nano-on-micro assemblies with a DNA/RNA synthesizer. A biotin-nanoparticle and immunomagnetic bead-based assay was developed. We succeeded in detecting Influenza A viruses directly in the lysis buffer supplemented with 10% saliva to simulate the clinical context. The biotin-nanoparticle amplification step enabled detection limits as low as 3 × 10 3 PFU mL −1 and 4 × 10 4 PFU mL −1 to be achieved for the H1N1 and H3N2 strains respectively. In contrast, a classical ELISA test based on the same antibody sandwich showed detection limit of 1.2 × 10 7 PFU mL −1 for H1N1. The new enhanced MELISA proved to be specific, as no cross-reactivity was found with a porcine respiratory virus (PRRSV).

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“…7,8 Solid phase synthesis (SPS) was recently extended as a bottom-up approach for efficient and modular functionalization of silica nanoparticles. 9 This method provides many advantages including high surface coverage and easy multifunctionalization, control of molecular orientation, and limitation of nonspecific adsorption. A wide range of molecules is available as phosphoramidite derivatives and thus can be directly incorporated on NP by SPS.…”

Section: ■ Introductionmentioning

confidence: 99%

Granulocyte Colony-Stimulating Factor Nanocarriers for Stimulation of the Immune System (Part I): Synthesis and Biodistribution Studies

et al. 2017

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In the field of cancer immunotherapy, an original approach consists of using granulocyte colony-stimulating factor (G-CSF) to target and activate neutrophils, cells of the innate immune system. G-CSF is a leukocyte stimulating molecule which is commonly used in cancer patients to prevent or reduce neutropenia. We focused herein on developing a G-CSF nanocarrier which could increase the in vivo circulation time of this cytokine, keeping it active for targeting the spleen, an important reservoir of neutrophils. G-CSF-functionalized silica and gold nanoparticles were developed. Silica nanoparticles of 50 nm diameter were functionalized by a solid phase synthesis approach. The technology enabled us to incorporate multiple functionalities on the surface such as a PEG as hydrophilic polymer, DTPA as In chelating agent and G-CSF. The gold nanocarrier consisted of nanoparticles of 2-3 nm diameter elaborated with DTPA groups on the surface and functionalized with G-CSF. We studied the particle biodistribution in mice with special attention to organs involved in the immune system. The two nanocarriers with similar functionalization of surface showed different pathways in mice, probably due to their difference in size. Considering the biodistribution after G-CSF functionalization, we confirmed that the protein was capable of modifying the pharmacokinetics by increasing the nanocarrier concentration in the spleen, a reservoir of G-CSF receptor expressing cells.

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Oligonucleotide solid-phase synthesis on fluorescent nanoparticles grafted on controlled pore glass

Cited by 9 publications

References 32 publications

Highly Sensitive Detection ofCampylobacter spp.in Chicken Meat using a Silica Nanoparticle Enhanced Dot Blot DNA Biosensor

Highly Sensitive Detection ofCampylobacter spp.in Chicken Meat using a Silica Nanoparticle Enhanced Dot Blot DNA Biosensor

Specific and sensitive detection of Influenza A virus using a biotin-coated nanoparticle enhanced immunomagnetic assay

Granulocyte Colony-Stimulating Factor Nanocarriers for Stimulation of the Immune System (Part I): Synthesis and Biodistribution Studies

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