Boronate affinity nanoparticles for RNA isolation

Toprak, Aykut; Görgün, Cansu; Kuru, Cansu İlke; Türkcan, Ceren; Uygun, Murat; Akgöl, Sinan

doi:10.1016/j.msec.2014.11.033

Cited by 23 publications

(10 citation statements)

References 22 publications

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“…The maximum adsorption capacity of the poly(HEMA‐GuaM)‐100 cryogels was found as 11.94 mg RNA/g poly(HEMA‐GuaM)‐100 cryogel. This value is quite high as compared to those of in the literature .…”

Section: Resultsmentioning

confidence: 44%

PolyGuanine methacrylate cryogels for ribonucleic acid purification

Köse

Uzun

2016

J of Separation Science

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The isolation and purification of ribonucleic acid have attracted attention recently for the understanding of the functions in detail because of the necessity for the treatment of genetic diseases. In this study, guanine-incorporated polymeric cryogels were developed to obtain highly purified ribonucleic acid. The satisfactory purification performance was achieved with the guanine-incorporated poly (2-hydroxyethyl methacrylate-guanine methacrylate) cryogels. The most crucial advantages to use guanine as a functional monomer are to obtain a real natural interaction between guanine on the polymeric material and cytosine on the ribonucleic acid. Moreover, using cryogel with a highly porous structure and high swelling ratio provide advantages of getting more water within the structure to get more analyte to interact. The characterization of cryogels has proved the success of the synthesis and the perfect natural interaction to be taken place between the ligand (guanine methacrylate) and the cytosine in the ribonucleic acid molecules. Although the pores within the structure of cryogels are small, they provide efficient and fast adsorption. The chromatographic separation performance was investigated for different conditions (pH, temperature etc.). The desorption ratio and reusability were also analyzed at the end of the five adsorption-desorption cycles with no significant changes.

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Section: Resultsmentioning

confidence: 44%

PolyGuanine methacrylate cryogels for ribonucleic acid purification

Köse

Uzun

2016

J of Separation Science

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“…Surface functionalization of nanoparticles is not limited to chemicals and synthetic compounds, but rather a large aim of biomedical nanotechnology focuses on the loading of biomolecules including protein, DNA, and RNA onto nanoparticles 32 33 34 . Typically RNA cannot be detected by fluorescence spectroscopy without the aid of a dye, however, 2D FDS can further be used to detect successful RNA adsorption to the nanoparticle surface.…”

Section: Resultsmentioning

confidence: 99%

Two-Dimensional Fluorescence Difference Spectroscopy to Characterize Nanoparticles and their Interactions

Hurst

Delong

2016

Sci Rep

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Two dimensional fluorescence difference spectroscopy (2D FDS) detects nanoparticle interactions following surface functionalization and biomolecule loading by generating a spectral signature of the fluorescent intensity per excitation and emission wavelengths. Comparing metal oxide nanoparticles revealed a unique spectral signature per material composition. 2D FDS showed to be sensitive to changes in surface properties between ZnO NPs synthesized by different methods. ZnO NP loaded with glycol chitosan, polyacrylic acid (PAA), or methoxy polyethylene glycol (mPEG) exhibited a distinct spectral signature shift. ZnO NP loaded with Torula Yeast RNA (TYRNA)(640 nm), polyinosinic: polycytidylic acid (pIC)(680 nm), or splice switching oligonucleotide (SSO)(650 nm) each revealed a shift in emission. Ras-Binding domain (RBD) at three concentrations (25, 37.5, 50 μg/mL) showed that fluorescent intensity was inversely related to the concentration of protein loaded. These data support 2D FDS as a novel technique in identifying nanoparticles and their surface interactions as a quality assurance tool.

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“…In the adsorption analysis, the aniline ligand exhibited a much lower affinity than APBA ligand with boronic acid, implying that the boronic acid was very important in the binding mechanism. The other possible performance is for the boron atom to coordinate with the water molecule through intermolecular B-N coordination [ 34 ]. The Ser176 residue was located in the bottom of the active-site pocket that had enough space for binding with a molecule larger than the molecule of water.…”

Section: Resultsmentioning

confidence: 99%

“…Our preliminary virtual screening and experimental verification indicated that boronic acid derivatives (BADs) could reversibly inhibit the activity of MP [ 30 ]. Phenylboronate group, which can form a temporary covalent bond with any molecule that contains a 1,2- cis -diol group, is widely used in the affinity purification of 1,2- cis -diol-containing biomolecules such as glycoproteins, glycopeptides, nucleosides, and nucleic acids [ 31 , 32 , 33 , 34 , 35 ]. However, application of the resins modified by phenylboronate in the purification of metalloproteases has never been reported.…”

Section: Introductionmentioning

confidence: 99%

Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification

Wang²,

Xu³

et al. 2016

Marine Drugs

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Metalloproteases are emerging as useful agents in the treatment of many diseases including arthritis, cancer, cardiovascular diseases, and fibrosis. Studies that could shed light on the metalloprotease pharmaceutical applications require the pure enzyme. Here, we reported the structure-based design and synthesis of the affinity medium for the efficient purification of metalloprotease using the 4-aminophenylboronic acid (4-APBA) as affinity ligand, which was coupled with Sepharose 6B via cyanuric chloride as spacer. The molecular docking analysis showed that the boron atom was interacting with the hydroxyl group of Ser176 residue, whereas the hydroxyl group of the boronic moiety is oriented toward Leu175 and His177 residues. In addition to the covalent bond between the boron atom and hydroxyl group of Ser176, the spacer between boronic acid derivatives and medium beads contributes to the formation of an enzyme-medium complex. With this synthesized medium, we developed and optimized a one-step purification procedure and applied it for the affinity purification of metalloproteases from three commercial enzyme products. The native metalloproteases were purified to high homogeneity with more than 95% purity. The novel purification method developed in this work provides new opportunities for scientific, industrial and pharmaceutical projects.

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Boronate affinity nanoparticles for RNA isolation

Cited by 23 publications

References 22 publications

PolyGuanine methacrylate cryogels for ribonucleic acid purification

PolyGuanine methacrylate cryogels for ribonucleic acid purification

Two-Dimensional Fluorescence Difference Spectroscopy to Characterize Nanoparticles and their Interactions

Structure-Based Design and Synthesis of a New Phenylboronic-Modified Affinity Medium for Metalloprotease Purification

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