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https://www.readbyqxmd.com/read/28324833/simplified-purification-approach-of-urinary-neutrophil-gelatinase-associated-lipocalin-by-tangential-flow-filtration-and-ion-exchange-chromatography
#1
Kunal K Shukla, Shamkant B Badgujar, Paresh B Bhanushali, Sushma G Sabharwal
This investigation reports a simplified approach for the purification of urinary siderocalin known as neutrophil gelatinase-associated lipocalin (NGAL). Urinary NGAL was purified by tangential flow filtration and ion exchange chromatography. Isolated NGAL was analyzed by SDS-PAGE, immunoblotting and mass spectrometry (MS). The relative molecular mass of NGAL is 23674Da. Peptide mass fingerprinting of the purified NGAL yielded peptides that partially matched with known sequence of P80188 (NGAL_HUMAN). The tryptic digestion profile of isolated NGAL infers that it may be unique and additive molecule in the dictionary of urinary proteins...
March 11, 2017: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
https://www.readbyqxmd.com/read/28324719/negatively-charged-polysulfone-membranes-with-hydrophilicity-and-antifouling-properties-based-on-in-situ-cross-linked-polymerization
#2
Lijing Zhu, Haiming Song, Dawei Zhang, Gang Wang, Zhixiang Zeng, Qunji Xue
Polysulfone (PSf) membrane has been widely used in water separation and purification, although, membrane fouling is still a serious problem limiting its potential. We aim to improve the antifouling of PSf membranes via a very simple and efficient method. In this work, antifouling PSf membranes were fabricated via in situ cross-linked polymerization coupled with non-solvent induced phase separation. In brief, acrylic acid (AA) and vinyltriethoxysilane (VTEOS) were copolymerized in PSf solution, then directly casted into membranes without purification...
March 14, 2017: Journal of Colloid and Interface Science
https://www.readbyqxmd.com/read/28324713/monolithic-cobalt-doped-carbon-aerogel-for-efficient-catalytic-activation-of-peroxymonosulfate-in-water
#3
Peidong Hu, Mingce Long, Xue Bai, Cheng Wang, Caiyun Cai, Jiajun Fu, Baoxue Zhou, Yongfeng Zhou
As an emerging carbonaceous material, carbon aerogels (CAs) display a great potential in environmental cleanup. In this study, a macroscopic three-dimensional monolithic cobalt-doped carbon aerogel was developed by co-condensation of graphene oxide sheets and resorcinol-formaldehyde resin in the presence of cobalt ions, followed by lyophilization, carbonization and thermal treatment in air. Cobalt ions were introduced as a polymerization catalyst to bridge the organogel framework, and finally cobalt species were retained as both metallic cobalt and Co3O4, wrapped by graphitized carbon layers...
March 6, 2017: Journal of Hazardous Materials
https://www.readbyqxmd.com/read/28323416/one-pot-two-nanoprobe-assay-uncovers-targeted-glycoprotein-biosignature
#4
Mira Anne C Dela Rosa, Wei-Chun Chen, Yi-Ju Chen, Rofeamor P Obena, Chih-Hsiang Chang, Rey Y Capangpangan, Tung-Hung Su, Chi-Ling Chen, Pei-Jer Chen, Yu-Ju Chen
We report a one-pot two-nanoprobe approach coupled to mass spectrometry for simultaneous quantification and post-translational modification (PTM) profiling of targeted protein in biofluid. Using N-glycoprotein as model, the assay employs two nanoprobes, antibody-conjugated SiO2 nanoparticles and lectin-conjugated magnetic Fe3O4 nanoparticles, to achieve target glycoprotein isolation from biofluid and subsequent glycopeptide enrichment in a single tube. As demonstrated on α-fetoprotein (AFP), a serum biomarker for hepatocellular carcinoma (HCC), the assay has high purification specificity (20 glycopeptides) with 2-fold and 10-fold superior total glycopeptide intensity compared to non-one-pot method (9 glycopeptides) or without enrichment (6 glycopeptides), respectively...
March 21, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28319379/immobilization-of-%C3%AE-galactosidases-from-lactobacillus-on-chitin-using-chitin-binding-domain
#5
Mai Lan Mai Pham, Tatjana Leister, Hoang Anh Nguyen, Bien-Cuong Do, Tuan-Anh Pham, Dietmar Haltrich, Montarop Yamabhai, Thu-Ha Nguyen, Tien-Thanh Nguyen
Two β-galactosidases from Lactobacillus including a heterodimeric, LacLM type enzyme from L. reuteri L103 and a homodimeric LacZ type β-galactosidase from L. bulgaricus DSM 20081 were studied for immobilization on chitin using a carbohydrate-binding domain (chitin-binding domain, ChBD) from a chitinolytic enzyme. Three recombinant enzymes, namely LacLM-ChBD, ChBD-LacLM and LacZ-ChBD, were constructed and successfully expressed in L. plantarum WCFS1. Depending on the structure of the enzymes, either homodimeric or heterodimeric, as well as the positioning of the chitin-binding domain in relation to the catalytic domains, i...
March 20, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28319173/affinity-purification-of-human-m-calpain-through-an-intrinsically-disordered-inhibitor-calpastatin
#6
Hung Huy Nguyen, Mihaly Varadi, Peter Tompa, Kris Pauwels
Calpains are calcium-activated proteases that have biomedical and biotechnological potential. Their activity is tightly regulated by their endogenous inhibitor, calpastatin that binds to the enzyme only in the presence of calcium. Conventional approaches to purify calpain comprise multiple chromatographic steps, and are labor-intensive, leading to low yields. Here we report a new purification procedure for the human m-calpain based on its reversible calcium-mediated interaction with the intrinsically disordered calpastatin...
2017: PloS One
https://www.readbyqxmd.com/read/28319022/recovery-of-lignin-peroxidase-from-submerged-liquid-fermentation-of-amauroderma-rugosum-blume-t-nees-torrend-using-polyethylene-glycol-salt-aqueous-two-phase-system
#7
Wan Yng Linda Jong, Pau Loke Show, Tau Chuan Ling, Yee Shin Tan
Amauroderma rugosum is a wild mushroom species widely distributed in tropics and is classified under the class of Basidiomycetes. Basidiomycetes are well-known for their abilities of producing lignocellulolytic enzymes such as lignin peroxidase (LiP), laccase (Lac) and manganese peroxidase (MnP). Different factors such as nutrient sources, incubation period and agitation affect the production of lignocellulolytic enzymes. The A. rugosum produced LiP in the medium supplemented with potato dextrose broth (PDB), 0...
March 16, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28319020/in%C3%A2-vitro-preparation-and-assessment-of-radical-reducing-peptide-from-octopus-aegina-using-digestive-proteases
#8
Sekar Sudhakar, Rasool Abdul Nazeer
Antioxidant peptides protect biological macromolecules against radical damages. The use of these peptides was evaluated using free radicals scavenging assays [2,2-diphenyl-1 picrylhydrazyl (DPPH) and hydroxyl] with the help of UV-visible and electron spin resonance (ESR) spectroscopy methods. The Octopus aegina mantle protein were tested upon hydrolysis using gastrointestinal enzymes up to 12 h, where pepsin hydrolysate exhibited superior properties (DPPH: 44.39±0.67% and hydroxyl: 38.84±1.07%) compared with trypsin and α-chymotrypsin...
March 16, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28318569/designing-affinity-chromatographic-processes-for-the-capture-of-antibodies-part-i-a-simplified-approach
#9
D Pfister, L David, M Holzer, R-M Nicoud
Protein A affinity chromatography is a standard technique for the purification of therapeutic antibodies. Recently, multi-column chromatographic processes have emerged to turn classical capture processes into more efficient and continuous systems. The design of such chromatographic processes, be they single-column or multi-column systems, is described in this work. A rational method to conceive, scale-up and compare processes is proposed and illustrated with different examples. All along this article, the results of the equilibrium theory, i...
March 2, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28318567/determination-of-ochratoxin-a-in-wine-by-packed-in-tube-solid-phase-microextraction-followed-by-high-performance-liquid-chromatography-coupled-to-tandem-mass-spectrometry
#10
Mariane A Andrade, Fernando M Lanças
Ochratoxin A (OTA), a widely studied mycotoxin, can be found in a variety of food matrices. As its concentration in food is generally low (in the order of μg kg(-1)), sample preparation techniques are necessary for the analyte purification and pre-concentration in order to achieve the required low detection limits. The separation and detection methods used for OTA analysis should also offer proper sensitivity in order to allow the adequate quantification of the analyte. This manuscript addresses the development of a methodology aiming the analysis of OTA in wine samples by packed in-tube SPME in flow through extraction mode coupled to HPLC-MS/MS...
March 1, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28318247/metalized-nanocellulose-composites-as-a-feasible-material-for-membrane-supports-design-and-applications-for-water-treatment
#11
Perla Cruz-Tato, Edwin O Ortiz-Quiles, Karlene Vega-Figueroa, Liz Noemi Santiago-Martoral, Michael Flynn, Liz M Diaz-Vazquez, Eduardo Nicolau
Herein, we study the feasibility of using nanocellulose (NC)-based composites with silver and platinum nanoparticles as additive materials to fabricate the support layer of thin film composite (TFC) membranes for water purification applications. In brief, the NC surface was chemically modified and then was decorated with silver and platinum nanoparticles, respectively by chemical reduction. These metalized nanocellulose composites (MNC) were characterized by several techniques including: FTIR, XPS, TGA, XRD and XANES to probe their integrity...
March 20, 2017: Environmental Science & Technology
https://www.readbyqxmd.com/read/28317995/in-situ-deprotection-and-dynamic-covalent-assembly-using-a-dual-role-catalyst
#12
T Wei, J C Furgal, T F Scott
Utilization of constituent molecular precursors bearing covalently coreactive functional groups for self-assembly risks premature reaction, impeding synthetic and purification efforts. To prevent premature amine-aldehyde condensation for oligomers bearing both groups, we employ a dual-role Lewis acid catalyst for both in situ acetal deprotection and subsequent imine exchange, effecting oligomer assembly.
March 20, 2017: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/28317774/development-of-an-up-grading-process-to-produce-mlm-structured-lipids-from-sardine-discards
#13
R Morales-Medina, M Munio, A Guadix, E M Guadix
The aim of the work was to produce MLM structured lipids with caprylic acid (M) as medium chain fatty acid located at the external bonds of the glycerol backbone and concentrated polyunsaturated fatty acids (L) from sardine discards (Sardine pilchardus) in the central bond of the glycerol. To that end, the following steps were conducted: (i) fish oil extraction, (ii) Omega-3 free fatty acids (FFA) concentration (low temperature winterization), (iii) two-steps enzymatic esterification and (iv) triacylglycerols (TAG) purification (liquid column chromatography)...
August 1, 2017: Food Chemistry
https://www.readbyqxmd.com/read/28317743/synthesis-of-1-3-distearoyl-2-oleoylglycerol-by-enzymatic-acidolysis-in-a-solvent-free-system
#14
Xiaosan Wang, Yang Chen, Liyou Zheng, Qingzhe Jin, Xingguo Wang
1,3-Distearoyl-2-oleoylglycerol (SOS) is widely used as a cocoa butter improver and an anti-blooming agent in the chocolate industry. In this study, an effective process was developed to prepare SOS by enzymatic acidolysis. Under optimal reaction conditions (substrate molar ratio of 12, NS40086 loading of 10%, a solvent-free system, 75°C for 4h), a SOS yield of 70.2% was obtained. Subsequently, two-step purification was applied to purify the crude SOS product. Free fatty acids were completely removed after molecular distillation...
August 1, 2017: Food Chemistry
https://www.readbyqxmd.com/read/28317180/anti-acsa-2-defines-a-novel-monoclonal-antibody-for-prospective-isolation-of-living-neonatal-and-adult-astrocytes
#15
Christina G Kantzer, Camille Boutin, Ina D Herzig, Carolina Wittwer, Sandy Reiß, Marie Catherine Tiveron, Jan Drewes, Thomas D Rockel, Stefanie Ohlig, Jovica Ninkovic, Harold Cremer, Sandra Pennartz, Melanie Jungblut, Andreas Bosio
Astrocytes are the most abundant cell type of the central nervous system and cover a broad range of functionalities. We report here the generation of a novel monoclonal antibody, anti-astrocyte cell surface antigen-2 (Anti-ACSA-2). Flow cytometry, immunohistochemistry and immunocytochemistry revealed that Anti-ACSA-2 reacted specifically with a not yet identified glycosylated surface molecule of murine astrocytes at all developmental stages. It did not show any labeling of non-astroglial cells such as neurons, oligodendrocytes, NG2(+) cells, microglia, endothelial cells, leukocytes, or erythrocytes...
March 20, 2017: Glia
https://www.readbyqxmd.com/read/28316235/evaluation-of-nanobody-conjugates-and-protein-fusions-as-bioanalytical-reagents
#16
Virginia J Bruce, Brian R McNaughton
Enzyme-linked immunosorbent assay (ELISA), flow cytometry, and Western blot are common bioanalytical techniques. Successful execution traditionally requires the use of one or more commercially available antibody-small-molecule dye, or antibody-reporter protein conjugates that recognize relatively short peptide tags (<15 amino acids). However, the size of antibodies, and their molecular complexity (by virtue of post-translational disulfide formation and glycosylation) typically requires either expression in mammalian cells or purification from immunized mammals...
March 20, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28316046/treatment-of-municipal-wastewater-in-full-scale-on-site-sand-filter-reduces-bod-efficiently-but-does-not-reach-requirements-for-nitrogen-and-phosphorus-removal
#17
Petteri Laaksonen, Aki Sinkkonen, Gennadi Zaitsev, Esa Mäkinen, Timo Grönroos, Martin Romantschuk
A traditional sand filter for treatment of household wastewater was constructed in the fall of 2012 at Biolinja 12, Turku, Finland. Construction work was led and monitored by an authorized wastewater treatment consultant. The filter was placed on a field bordered by open ditches from all sides in order to collect excess rain and snowmelt waters. The filter was constructed and insulated from the environment so that all outflowing water was accounted for. Untreated, mainly municipal, wastewater from Varissuo suburb was pumped from a sewer separately via three septic tanks (volume = 1 m(3) each) into the filters...
March 18, 2017: Environmental Science and Pollution Research International
https://www.readbyqxmd.com/read/28316005/purification-kinetic-and-thermodynamic-characteristics-of-an-exo-polygalacturonase-from-penicillium-notatum-with-industrial-perspective
#18
Faiza Amin, Haq Nawaz Bhatti, Muhammad Bilal, Muhammad Asgher
An extracellular exo-polygalacturonase (exo-PG) produced by Penicillium notatum was purified (3.07-folds) by ammonium sulfate fractionation, ion exchange, and gel filtration chromatography. Two distinct isoforms of the enzyme, namely exo-PGI and exo-PGII, were identified during column purification with molecular weights of 85 and 20 kDa, respectively, on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme displayed its optimum activity at pH 6.0 and 50 °C and was found to be stable in the slightly acidic pH (ranging from 4...
March 18, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28315746/optimized-protocol-for-soluble-prokaryotic-expression-purification-and-structural-analysis-of-human-placenta-specific-1-plac1
#19
Mahboobeh Nazari, Amir-Hassan Zarnani, Roya Ghods, Rahman Emamzadeh, Somayeh Najafzadeh, Arash Minai-Tehrani, Jafar Mahmoudian, Maryam Yousefi, Sedigheh Vafaei, Sam Massahi, Mohammad-Reza Nejadmoghaddam
Placenta specific -1 (PLAC1) has been recently introduced as a small membrane-associated protein mainly involved in placental development. Expression of PLAC1 transcript has been documented in almost one hundred cancer cell lines standing for fourteen distinct cancer types. The presence of two disulfide bridges makes difficult to produce functional recombinant PLAC1 in soluble form with high yield. This limitation also complicates the structural studies of PLAC1, which is important for prediction of its physiological roles...
March 15, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28315745/production-of-low-expressing-recombinant-cationic-biopolymers-with-high-purity
#20
Xuguang Chen, Alireza Nomani, Niket Patel, Arash Hatefi
The growing complexity of recombinant biopolymers for delivery of bioactive agents requires the ability to control the biomaterial structure with high degree of precision. Genetic engineering techniques have provided this opportunity to synthesize biomaterials in an organism such as E. coli with full control over their lengths and sequences. One class of such biopolymers is recombinant cationic biopolymers with applications in gene delivery, regenerative medicine and variety of other biomedical applications...
March 15, 2017: Protein Expression and Purification
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