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https://www.readbyqxmd.com/read/28734606/thiol-yne-click-synthesis-of-boronic-acid-functionalized-silica-nanoparticle-graphene-oxide-composites-for-highly-selective-enrichment-of-glycoproteins
#1
Jingai Yang, Xiwen He, Langxing Chen, Yukui Zhang
In this work, a facile novel strategy is developed for the preparation of 4-mercapto-phenylboronic acid functionalized silica nanoparticle-graphene oxide composite (GO@MPBA) via thiol-yne click reaction. The morphology, structure and composition of the resulting GO@MPBA was characterized by transmission electron microscopy, X-ray powder diffraction, Fourier transform infrared spectroscopy, Fourier transform Raman spectroscopy, Thermogravimetric analysis and X-ray photoelectron spectrometry. GO@MPBA exhibited the high binding capacity towards glycoproteins such as ovalbumin (1288...
July 13, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28731680/nitrilotriacetic-amine-functionalized-polymeric-core-shell-nanoparticles-as-enzyme-immobilization-supports
#2
Dominic Keller, Ana Beloqui, Monica Martínez-Martínez, Manuel Ferrer, Guillaume Delaittre
Nitrilotriacetic amine (NTA) functionalized nanoparticles obtained by aqueous polymerization-induced self-assembly (PISA) are introduced as immobilization supports for polyhistidine-functionalized (His-tagged) enzymes. A novel initiator for nitroxide-mediated polymerization based on the nitroxide SG1 and carrying a protected NTA moiety was first synthesized. Size-exclusion chromatography (SEC) and electrospray ionization mass spectrometry (ESI-MS) proved the ability of this initiator to produce well-defined end-functional vinyl polymers...
July 21, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/28729217/preventing-postoperative-tissue-adhesion-using-injectable-carboxymethyl-cellulose-pullulan-hydrogels
#3
Sumi Bang, Young-Gwang Ko, Won Il Kim, Donghwan Cho, Won Ho Park, Oh Hyeong Kwon
An injectable adhesive hydrogel composed of carboxymethyl cellulose (CMC) and pullulan is developed and evaluated as a postoperative anti-adhesion barrier. CMC was modified with tyramine to introduce crosslinking via an EDC-NHS reaction. The in situ hydrogel was developed by an enzyme-mediated reaction of tyramine-immobilized CMC with horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). Pullulan was added to the hydrogel solution to improve adhesiveness to the wound area and accelerate biodegradation...
July 17, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28728930/specific-enrichment-of-glycoproteins-with-polymer-monolith-functionalized-with-glycocluster-grafted-%C3%AE-cyclodextrin
#4
Hai-Jiao Zheng, Jiu-Tong Ma, Wei Feng, Qiong Jia
The low abundance of glycoproteins in complex samples results in the prerequisite role of efficient and selective enrichment of them. In the present work, we designed a new kind of glycosylation poly(hydroxyethyl methacrylate-pentaerythritol triacrylate) monolith functionalized with glycocluster grafted β-cyclodextrin for the enrichment of glycoproteins. The introduced modifiers endowed the monolithic material with enhanced hydrophilicity and surface area, which benefitted to improve the enrichment selectivity and extraction efficiency for glycopeptides...
July 11, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28726378/ptnps-as-scaffolds-to-regulate-interenzyme-distance-for-construction-of-efficient-enzyme-cascade-amplification-for-ultrasensitive-electrochemical-detection-of-mmp-2
#5
Beibei Kou, Yaqin Chai, Yali Yuan, Ruo Yuan
The high catalytic efficiency of enzyme cascade reaction mainly depends on optimal interenzyme distance regulated by the special scaffolds. In this work, the rigid PtNPs with different sizes were employed as scaffolds to regulate interenzyme distance for efficient enzyme cascade amplification to construct electrochemical biosensor for sensitive detection of matrix metalloproteinases-2 (MMP-2), which overcame the drawbacks of instable construction and sophisticated preparation induced by conventional scaffolds such as metal-organic frameworks (MOFs), DNA nanostructures...
July 20, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28722272/development-of-a-thiol-ene-based-screening-platform-for-enzyme-immobilization-demonstrated-using-horseradish-peroxidase
#6
Christian Hoffmann, Manuel Pinelo, John M Woodley, Anders E Daugaard
Efficient immobilization of enzymes on support surfaces requires an exact match between the surface chemistry and the specific enzyme. A successful match would normally be identified through time consuming screening of conventional resins in multiple experiments testing individual immobilization strategies. In this study we present a versatile strategy that largely expands the number of possible surface functionalities for enzyme immobilization in a single, generic platform. The combination of many individual surface chemistries and thus immobilization methods in one modular system permits faster and more efficient screening, which we believe will result in a higher chance of discovery of optimal surface/enzyme interactions...
July 19, 2017: Biotechnology Progress
https://www.readbyqxmd.com/read/28721200/protein-bound-polyphenols-create-ghost-band-artifacts-during-chemiluminescence-based-antigen-detection
#7
Nathalie Plundrich, Mary Ann Lila, Edward Foegeding, Scott Laster
Antigen detection during Western blotting commonly utilizes a horseradish peroxidase-coupled secondary antibody and enhanced chemiluminescent substrate. We utilized this technique to examine the impact of green tea-derived polyphenols on the binding of egg white protein-specific IgE antibodies from allergic human plasma to their cognate antigens. Our experiments unexpectedly showed that green tea-derived polyphenols, when stably complexed with egg white proteins, caused hyperactivation of horseradish peroxidase resulting in the appearance of white "ghost" bands...
2017: F1000Research
https://www.readbyqxmd.com/read/28718841/development-of-an-immunosensor-for-pfhrp-2-as-a-biomarker-for-malaria-detection
#8
Aver Hemben, Jon Ashley, Ibtisam E Tothill
Plasmodium falciparum histidine-rich protein 2 (PfHRP 2) was selected in this work as the biomarker for the detection and diagnosis of malaria. An enzyme-linked immunosorbent assay (ELISA) was first developed to evaluate the immunoreagent's suitability for the sensor's development. A gold-based sensor with an integrated counter and an Ag/AgCl reference electrode was first selected and characterised and then used to develop the immunosensor for PfHRP 2, which enables a low cost, easy to use, and sensitive biosensor for malaria diagnosis...
July 18, 2017: Biosensors
https://www.readbyqxmd.com/read/28714899/electrochemical-flow-elisa-for-rapid-and-sensitive-determination-of-microcystin-lr-using-automated-sequential-injection-system
#9
Lesedi Lebogang, Jongjit Jantra, Martin Hedström, Bo Mattiasson
An amperometric immunoanalysis system based on monoclonal antibodies immobilized on Sepharose beads and packed into a micro-immunocolumn was developed for the quantification of microcystin-LR. Microcystin-LR (MCLR) was used as a reference microcystin variant. Inside the immunocolumn, free microcystins and microcystin-horseradish peroxidase (tracer) were sequentially captured by the immobilized antibodies, and the detection was performed electrochemically using Super AquaBlue ELISA substrate 2,2'-azinobis(3-ethylbenzothiazoline-sulfonic acid) (ABTS)...
July 16, 2017: Sensors
https://www.readbyqxmd.com/read/28712997/novel-characteristics-of-horseradish-peroxidase-immobilized-onto-the-polyvinyl-alcohol-alginate-beads-and-its-methyl-orange-degradation-potential
#10
Muhammad Bilal, Tahir Rasheed, Hafiz M N Iqbal, Hongbo Hu, Wei Wang, Xuehong Zhang
Herein, we report the immobilization of in-house isolated horseradish peroxidase (HRP) from Armoracia rusticana with novel characteristics. The HRP was immobilized onto the self-fabricated polyvinyl alcohol-alginate (PVA-alginate) beads using sodium nitrate as a cross-linker. The PVA-alginate beads (2.0mm size) developed using 10% PVA and 1.5% sodium alginate showed maximal immobilization yield. The surface morphologies of the PVA-alginate (control) and immobilized-HRP were characterized by scanning electron microscopy (SEM)...
July 13, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28712944/a-fe3o4-au-basedpseudo-homogeneous-electrochemical-immunosensor-for-afp-measurement-using-afp-antibody-gnps-hrp-as-detection-probe
#11
Yulin Yuan, Shanshan Li, Yewei Xue, Jintao Liang, Lijie Cui, Qingbo Li, Sufang Zhou, Yong Huang, Guiyin Li, Yongxiang Zhao
In this study, a Fe3O4@Au-based pseudo-homogeneous electrochemical immunosensor was prepared for detection of alpha fetoprotein (AFP), a well-known hepatocellular carcinoma biomarker. The primary antibody (Ab1) was immobilized on Fe3O4@Au NPs as the capture probe. Horseradish peroxidase (HRP) and secondary antibody (Ab2) were conjugated on gold nanoparticles (GNPs) through electrostatic adsorption to form signal-amplifying labels. In the presence of AFP, a sandwich immunocomplex was formed via specific recognition of antigen-antibody in a Fe3O4@Au-basedpseudo-homogeneousreaction system...
July 13, 2017: Analytical Biochemistry
https://www.readbyqxmd.com/read/28710930/immobilization-effects-on-the-photocatalytic-activity-of-cds-quantum-dots-horseradish-peroxidase-hybrid-nanomaterials
#12
Iker Iñarritu, Eduardo Torres, Antonio Topete, José Campos-Terán
The potential use of hybrid nanomaterials based on inorganic optically active nanoparticles known as quantum dots (QDs) and horseradish peroxidase (HRP) has been proposed by several authors as light-controllable nanocatalyzers, moreover, the immobilization within or over silica based supports represents an advantage over bulk-dispersed systems. However, the implications of the immobilization of such hybrid photoactivatable catalyzing systems have not been clarified with detail. Here, we present a thorough study of the functional photoactive efficiency and recycling of immobilized CdS QDs and HRP systems with different configurations, immobilized over silanized silica quartz crystal microbalance (QCM) sensors, allowing an accurate measure of the immobilized mass of each component and its correlation with the initial reaction rate of conversion of Amplex Red (AR) to resorufin...
July 6, 2017: Journal of Colloid and Interface Science
https://www.readbyqxmd.com/read/28709962/the-role-of-macrophages-in-the-development-of-biliary-injury-in-a-lipopolysaccharide-aggravated-hepatic-ischaemia-reperfusion-model
#13
REVIEW
J Reiling, K R Bridle, F G Schaap, L Jaskowski, N Santrampurwala, L J Britton, C M Campbell, P L M Jansen, S W M Olde Damink, D H G Crawford, C H C Dejong, J Fawcett
INTRODUCTION: Endotoxins, in the form of lipopolysaccharides (LPS), are potent inducers of biliary injury. However the mechanism by which injury develops remains unclear. We hypothesized that hepatic macrophages are pivotal in the development of endotoxin-induced biliary injury and that no injury would occur in their absence. MATERIAL AND METHODS: Clodronate liposomes were used to deplete macrophages from the liver. Forty-eight rats were equally divided across six study groups: sham operation (sham), liposome treatment and sham operation (liposomes+sham), 1mg/kg LPS i...
July 11, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28703007/a-chaperonin-nanocaged-hemin-as-an-artificial-metalloenzyme-for-oxidation-catalysis
#14
Xiaoqiang Wang, Chao Wang, Meihong Pan, Junting Wei, Fuping Jiang, Rongsheng Lu, Xuan Liu, Yihui Huang, Fang Huang
Taking inspiration from biology's effectiveness in functionalizing protein-based nanocages for chemical processes, we describe here a rational design of an artificial metalloenzyme for oxidations with the bacterial chaperonin GroEL, a nanocage for protein folding in nature, by supramolecular anchoring of catalytically active hemin in its hydrophobic central cavity. The promiscuity of the chaperonin cavity is an essential element of this design, which can mimic the hydrophobic binding pocket in natural metalloenzymes to accept cofactor and substrate without requiring specific ligand-protein interactions...
July 13, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28702124/6-hydroxy-5-nitrobenzo-d-oxazol-2-3h-one-a-degradable-derivative-of-natural-6-hydroxybenzoxazolin-2-3h-one-produced-by-pantoea-ananatis
#15
Margot Schulz, Dieter Sicker, Oliver Schackow, Lothar Hennig, Diana Hofmann, Ulrich Disko, Marina Ventura, Kateryna Basyuk
Pantoea ananatis is a bacterium associated with other microorganisms on Abutilon theophrasti Medik. roots. It converts 6-hydroxybenzoxazolin-2(3H)-one (BOA-6-OH), a hydroxylated derivative of the allelochemical benzoxazolin-2(3H)-one, into 6-hydroxy-5-nitrobenzo[d]oxazol-2(3H)-one. The compound was identified by NMR and mass spectrometric methods. In vitro synthesis succeeded with Pantoea protein, with isolated proteins from the Abutilon root surface or with horseradish peroxidase in the presence of nitrite and H2O2...
2017: Communicative & Integrative Biology
https://www.readbyqxmd.com/read/28697412/conformation-and-activity-alteration-of-horseradish-peroxidase-induced-by-the-interaction-with-gene-carrier-polyethyleneimines
#16
Aimin Huang, Bangzhi Wei, Junyong Mo, Yajing Wang, Lin Ma
Polyethyleneimine (PEI) has long been considered as "golden standard" for polymeric gene delivery carriers. However the molecular basis of the cytotoxicity of PEI is poorly understood. Little is known about the effects of PEI on the structure and functions of biomacromolecules. In this work, fluorescence, UV-vis absorption, circular dichroism spectroscopy were conducted to investigate the influence of PEI of average molecular weight 25, 10 and 1.8kDa (denoted as PEI25k, PEI10k and PEI1.8k) on the conformation of horseradish peroxidase (HRP) and its catalytic efficiency...
July 1, 2017: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
https://www.readbyqxmd.com/read/28696095/dt-diaphorase-as-a-bifunctional-enzyme-label-that-allows-rapid-enzymatic-amplification-and-electrochemical-redox-cycling
#17
Cheolho Kang, Juyeon Kang, Nam-Sihk Lee, Young Ho Yoon, Haesik Yang
The most common enzyme labels in enzyme-linked immunosorbent assays are alkaline phosphatase and horseradish peroxidase, which, however, have some limitations for use in electrochemical immunosensors. This Article reports that the small and thermostable DT-diaphorase (DT-D) and electrochemically inactive 4-nitroso-1-naphthol (4-NO-1-N) can be used as a bifunctional enzyme label and a rapidly reacting substrate, respectively, for electrochemical immunosensors. This enzyme-substrate combination allows high signal amplification via rapid enzymatic amplification and electrochemical redox cycling...
July 11, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28692846/development-of-screen-printed-electrode-based-immunosensor-for-the-detection-of-her2-antigen-in-human-serum-samples
#18
Sagarika Deepthy Tallapragada, Keya Layek, Runu Mukherjee, Kalyan Kumar Mistry, Monidipa Ghosh
In this study, an immunosensor based on screen-printed electrode (SPE) has been developed for the detection of Human Epidermal Growth Factor Receptor-2 (HER2) antigen. The SPEs were fabricated and a sandwich enzyme linked immunosorbent assay (ELISA) format was followed for the construction of the immunosensor. The capture antibody (mouse anti-human ErbB2) was coated onto the electrode surface without any prior surface modification, followed by the addition of recombinant human HER2 antigen. Biotinylated goat anti-human ErbB2 was used as the detection antibody which was linked to streptavidin conjugated horseradish peroxidase (HRP)...
June 30, 2017: Bioelectrochemistry
https://www.readbyqxmd.com/read/28690263/direct-electron-transfer-type-bioelectrocatalysis-of-peroxidase-at-mesoporous-carbon-electrodes-and-its-application-for-glucose-determination-based-on-bienzyme-system
#19
Hong-Qi Xia, Yuki Kitazumi, Osamu Shirai, Kenji Kano
Non-catalytic direct electron transfer (DET) signal of Compound I of horseradish peroxidase (POD) was first detected at 0.7 V on POD/carbon nanotube mixture-modified electrodes. Excellent performance of DET-type bioelectrocatalysis was achieved with POD immobilized with glutaraldehyde on Ketjen Black (KB)-modified electrodes for H2O2 reduction with an onset potential of 0.65 V (vs. Ag | AgCl | sat. KCl) without any electrode surface modification. The POD-immobilized KB electrode was found to be suitable for detecting H2O2 with a low detection limit (0...
2017: Analytical Sciences: the International Journal of the Japan Society for Analytical Chemistry
https://www.readbyqxmd.com/read/28685478/higher-fungi-are-a-rich-source-of-l-amino-acid-oxidases
#20
Gašper Žun, Janko Kos, Jerica Sabotič
L-Amino acid oxidases (LAO) are widely distributed enzymes but those from snake venoms have been studied the most. We describe a method for in-gel detection of LAO activities based on H2O2 detection by a horseradish peroxidase-coupled reaction using o-phenylenediamine. Complex substrates and single L-amino acids were used successfully for screening LAO activities in higher fungi using crude aqueous extracts of fruiting bodies of 22 basidiomycetes and 1 ascomycete. Half of these samples exhibited one to two bands of LAO activities with mostly broad substrate specificities and a variety of apparent molecular masses ranging from 25 to 200 kDa that were generally more active at pH 5...
July 2017: 3 Biotech
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