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Affinity chromatography

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https://www.readbyqxmd.com/read/28087367/production-of-recombinant-proteins-in-escherichia-coli-tagged-with-the-fusion-protein-cusf3h
#1
Teresa Vargas-Cortez, Jose Ruben Morones-Ramirez, Isaias Balderas-Renteria, Xristo Zarate
Recombinant protein expression in the bacterium Escherichia coli still is the number one choice for large-scale protein production. Nevertheless, many complications can arise using this microorganism, such as low yields, the formation of inclusion bodies, and the requirement for difficult purification steps. Most of these problems can be solved with the use of fusion proteins. Here, the use of the metal-binding protein CusF3H+ is described as a new fusion protein for recombinant protein expression and purification in E...
January 10, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28087106/charge-variant-analysis-of-proposed-biosimilar-to-trastuzumab
#2
Pravinkumar Dakshinamurthy, Pavithra Mukunda, Bhargav Prasad Kodaganti, Bharath Ravindra Shenoy, Bairavabalakumar Natarajan, Amol Maliwalave, Vivek Halan, Sathyabalan Murugesan, Sunit Maity
Trastuzumab is a humanized monoclonal antibody (mAb) employed for the treatment of HER2 Positive Breast Cancer. A HER2 overexpressing tumor cell binds to Trastuzumab and attracts immune cells which lead to induction of Antibody Dependent Cellular Cytotoxicity (ADCC) by binding to Fc receptors (CD16a or FcγRIIIa) on an effector cell, such as natural killer (NK) cells. The most commonly expressed receptor on NK cell is CD16a which binds to the Fc portion of Trastuzumab. The ligand-independent HER2-HER3 dimerization is the most potent stimulator of downstream pathways for regulation of cell growth and survival...
January 10, 2017: Biologicals: Journal of the International Association of Biological Standardization
https://www.readbyqxmd.com/read/28087055/graft-modification-of-cotton-with-phosphate-group-and-its-application-to-the-enrichment-of-phosphopeptides
#3
Xiao-Mei He, Xi Chen, Bi-Feng Yuan, Yu-Qi Feng
Grafting copolymerization, especially "grafting from" approach, has attracted a great attention for the preparation of cellulose-based materials with various functionalities. In this study, a novel phosphate group-containing cotton fiber-based material, CF-NH2-AZO-p(VPA-x), was successfully synthesized by a "grafting from" radical polymerization approach using vinyl phosphonic acid (VPA) as monomer. The phosphate group content of CF-NH2-AZO-p(VPA-x) could be easily regulated by adjusting the concentration of VPA monomer...
January 9, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28079114/discovering-potential-serological-biomarker-for-chronic-hepatitis-b-virus-related-hepatocellular-carcinoma-in-chinese-population-by-mal-associated-serum-glycoproteomics-analysis
#4
Tianhua Liu, Denghe Liu, Riqiang Liu, Hucong Jiang, Guoquan Yan, Wei Li, Lu Sun, Shu Zhang, Yinkun Liu, Kun Guo
The accuracy of current biomarkers for the diagnosis of hepatocellular carcinoma (HCC), especially chronic Hepatitis B Virus (HBV)-related HCC, is limited. Recent progress in glycoproteomics has provided a novel platform for screening novel serological biomarkers of HCC. In this study, lectin affinity chromatography by Maackia amurensis lectin (MAL) and iTRAQ combined with mass spectrometric analysis were performed to enrich and identify the glycoprotein fractions in serum samples from HBV-related HCC patients and from healthy controls...
January 12, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28076454/a-broad-ph-range-and-processive-chitinase-from-a-metagenome-library
#5
S S Thimoteo, A Glogauer, H Faoro, E M de Souza, L F Huergo, B M Moerschbacher, F O Pedrosa
Chitinases are hydrolases that degrade chitin, a polymer of N-acetylglucosamine linked β(1-4) present in the exoskeleton of crustaceans, insects, nematodes and fungal cell walls. A metagenome fosmid library from a wastewater-contaminated soil was functionally screened for chitinase activity leading to the isolation and identification of a chitinase gene named metachi18A. The metachi18A gene was subcloned and overexpressed in Escherichia coli BL21 and the MetaChi18A chitinase was purified by affinity chromatography as a 6xHis-tagged fusion protein...
January 5, 2017: Brazilian Journal of Medical and Biological Research, Revista Brasileira de Pesquisas Médicas e Biológicas
https://www.readbyqxmd.com/read/28075189/11-analytically-confirmed-cases-of-mexedrone-use-among-polydrug-users
#6
Liam Roberts, Loretta Ford, Neel Patel, J Allister Vale, Sally M Bradberry
INTRODUCTION: Mexedrone, 3-methoxy-2-(methylamino)-1-(4-methylphenyl)propan-1-one, is the alpha-methoxy-derivative of mephedrone (4-methyl-N-methyl cathinone). Mexedrone inhibits the re-uptake of serotonin and dopamine in a dose-dependent manner and has affinity for serotonin and dopamine membrane transporters and receptors (5-HT2 and D2 receptors), producing sympathomimetic effects similar to amfetamines. To date there are no published clinical reports on mexedrone use that are analytically confirmed...
January 11, 2017: Clinical Toxicology
https://www.readbyqxmd.com/read/28068756/hydrophilic-mesoporous-silica-materials-for-highly-specific-enrichment-of-n-linked-glycopeptide
#7
Nianrong Sun, Jiawen Wang, Jizong Yao, Chun-Hui Deng
Hydrophilic interaction liquid chromatography (HILIC) is a significant enrichment strategy in glycoproteomics profiling. In this report, hydrophilic magnetic mesoporous silica materials (Fe3O4@mSiO2-IDA) were designed and synthesized as an outstanding enrichment platform for glycopeptide analysis. By taking advantage of their merits, such as large surface area, excellent hydrophilicity and unbiased affinity toward all types of glycopeptides, the Fe3O4@mSiO2-IDA nanomaterials were successfully applied to capture glycopeptides from complex samples...
January 4, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28063296/biodegradation-of-the-artificial-sweetener-acesulfame-in-biological-wastewater-treatment-and-sandfilters
#8
Sandro Castronovo, Arne Wick, Marco Scheurer, Karsten Nödler, Manoj Schulz, Thomas A Ternes
A considerable removal of the artificial sweetener acesulfame (ACE) was observed during activated sludge processes at 13 wastewater treatment plants (WWTPs) as well as in a full-scale sand filter of a water works. A long-term sampling campaign over a period of almost two years revealed that ACE removal in WWTPs can be highly variable over time. Nitrifying/denitrifying sequencing batch reactors (SBR) as well as aerobic batch experiments with activated sludge and filter sand from a water works confirmed that both activated sludge as well as filter sand can efficiently remove ACE and that the removal can be attributed to biologically mediated degradation processes...
November 17, 2016: Water Research
https://www.readbyqxmd.com/read/28062799/a-two-pronged-binding-mechanism-of-igg-to-the-neonatal-fc-receptor-controls-complex-stability-and-igg-serum-half-life
#9
Pernille Foged Jensen, Angela Schoch, Vincent Larraillet, Maximiliane Hilger, Tilman Schlothauer, Thomas Emrich, Kasper Dyrbert Rand
The success of recombinant monoclonal immunoglobulins (IgG) is rooted in their ability to target distinct antigens with high affinity combined with an extraordinarily long serum half-life, typically around three weeks. The pharmacokinetics of IgGs is intimately linked to the recycling mechanism of the neonatal Fc receptor (FcRn). For long serum half-life of therapeutic IgGs, the highly pH dependent interaction with FcRn needs to be balanced to allow efficient FcRn binding and release at slightly acidic pH and physiological pH, respectively...
January 6, 2017: Molecular & Cellular Proteomics: MCP
https://www.readbyqxmd.com/read/28060902/preference-of-conjugated-bile-acids-over-unconjugated-bile-acids-as-substrates-for-oatp1b1-and-oatp1b3
#10
Takahiro Suga, Hiroaki Yamaguchi, Toshihiro Sato, Masamitsu Maekawa, Junichi Goto, Nariyasu Mano
Bile acids, the metabolites of cholesterol, are signaling molecules that play critical role in many physiological functions. They undergo enterohepatic circulation through various transporters expressed in intestine and liver. Human organic anion-transporting polypeptides (OATP) 1B1 and OATP1B3 contribute to hepatic uptake of bile acids such as taurocholic acid. However, the transport properties of individual bile acids are not well understood. Therefore, we selected HEK293 cells overexpressing OATP1B1 and OATP1B3 to evaluate the transport of five major human bile acids (cholic acid, chenodeoxycholic acid, deoxycholic acid, ursodeoxycholic acid, lithocholic acid) together withtheir glycine and taurine conjugates via OATP1B1 and OATP1B3...
2017: PloS One
https://www.readbyqxmd.com/read/28058522/identification-and-characterization-of-a-novel-carboxylesterase-fpbh-that-hydrolyzes-aryloxyphenoxypropionate-herbicides
#11
Chenghong Wang, Jiguo Qiu, Youjian Yang, Jinwei Zheng, Jian He, Shunpeng Li
OBJECTIVE: To identify and characterize a novel aryloxyphenoxypropionate (AOPP) herbicide-hydrolyzing carboxylesterase from Aquamicrobium sp. FPB-1. RESULTS: A carboxylesterase gene, fpbH, was cloned from Aquamicrobium sp. FPB-1. The gene is 798 bp long and encodes a protein of 265 amino acids. FpbH is smaller than previously reported AOPP herbicide-hydrolyzing carboxylesterases and shares only 21-35% sequence identity with them. FpbH was expressed in Escherichia coli BL21(DE3) and the product was purified by Ni-NTA affinity chromatography...
January 5, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28057569/a-novel-peroxidase-purified-from-marsdenia-megalantha-latex-inhibits-phytopathogenic-fungi-mediated-by-cell-membrane-permeabilization
#12
Henrique P Oliveira, Rodolpho G G Silva, Jose T A Oliveira, Daniele O B Sousa, Mirella L Pereira, Pedro F N Souza, Arlete A Soares, Valdirene M Gomes, Ana C O Monteiro-Moreira, Frederico B M B Moreno, Ilka M Vasconcelos
An antifungal class III peroxidase was purified from Marsdenia megalantha latex (named Mo-POX) using DEAE-cellulose and gel filtration chromatography on a Superose 12 HR 10/30 column. Mm-POX has an apparent molecular mass of 67.0kDa and a pI of 5.2, shares identity with other peroxidases, and follows Michaelis-Menten kinetics. It has a high affinity for guaiacol and hydrogen peroxide. The pH and temperature optima for Mm-POX were 5.0-7.0 and 60°C, respectively. The catalytic activity of Mm-POX was decreased in the presence of classic peroxidase inhibitors including azide, dithiothreitol, ethylenediamine tetraacetic acid, and sodium metabisulfite and high concentrations of Na(+), Mn(+), and salicylic acid...
January 3, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28056947/triptolide-induces-hepatotoxicity-via-inhibition-of-cyp450s-in-rat-liver-microsomes
#13
Yan Lu, Tong Xie, Yajie Zhang, Fuqiong Zhou, Jie Ruan, Weina Zhu, Huaxu Zhu, Zhe Feng, Xueping Zhou
BACKGROUND: Triptolide (TP), an active constituent of Tripterygium wilfordii, possesses numerous pharmacological activities. However, its effects on cytochrome P450 enzymes (CYP450s) in rats remain unexplored. METHODS: In this study, the effects of triptolide on the six main CYP450 isoforms (1A2, 2C9, 2C19, 2D6, 2E1, and 3A) were investigated both in vivo and in vitro. We monitored the body weight, survival proportions, liver index, changes in pathology, and biochemical index upon TP administration, in vivo...
January 5, 2017: BMC Complementary and Alternative Medicine
https://www.readbyqxmd.com/read/28054967/expression-purification-and-biophysical-characterization-of-a-secreted-anthrax-decoy-fusion-protein-in-nicotiana-benthamiana
#14
Kalimuthu Karuppanan, Sifti Duhra-Gill, Muchena J Kailemia, My L Phu, Carlito B Lebrilla, Abhaya M Dandekar, Raymond L Rodriguez, Somen Nandi, Karen A McDonald
Anthrax toxin receptor-mediated drug development for blocking anthrax toxin action has received much attention in recent decades. In this study, we produced a secreted anthrax decoy fusion protein comprised of a portion of the human capillary morphogenesis gene-2 (CMG2) protein fused via a linker to the fragment crystallizable (Fc) domain of human immunoglobulin G1 in Nicotiana benthamiana plants using a transient expression system. Using the Cauliflower Mosaic Virus (CaMV) 35S promoter and co-expression with the p19 gene silencing suppressor, we were able to achieve a high level of recombinant CMG2-Fc-Apo (rCMG2-Fc-Apo) protein accumulation...
January 4, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28049027/an-integrated-shotgun-proteomics-and-bioinformatics-approach-for-analysis-of-brain-proteins-from-mcao-model-using-serial-affinity-chromatograph-with-four-active-ingredients-from-shengmai-preparations-as-ligands
#15
Xiao-Xue Fan, Fang Li, Yan-Ni Lv, Yu Zhang, Jun-Ping Kou, Bo-Yang Yu
Identification and validation of disease-relevant target proteins for natural products is an essential component of modern pharmaceutical research. In the present study, an integrated shotgun proteomics and bioinformatics approach was established to profile the interaction of active small molecules derived from ShengMai preparations (SMXZF) with hundreds of endogenously expressed proteins from middle cerebral artery occlusion (MCAO) model. Affinity-based proteomic strategies for isolation and identification of targets for the bioactive components is a classic, but still powerful approach...
January 1, 2017: Neurochemistry International
https://www.readbyqxmd.com/read/28043508/proteome-scale-mdr-tb-antibody-responses-for-identification-of-putative-biomarkers-for-the-diagnosis-of-drug-resistant-mycobacterium-tuberculosis
#16
Shamsi Yari, Alireza Hadizadeh Tasbiti, Mostafa Ghanei, Seyed Davar Siadat, Fatemeh Yari, A Bahrmand
OBJECTIVE: Multidrug-resistant tuberculosis (MDR-TB) is caused by Mycobacterium tuberculosis strains that do not respond to isoniazid and rifampicin, the two most effective first-line anti-TB drugs. Here, we designed and produced antibodies based on biomarkers that exist only in MDR-TB. METHODS: Bacilli were cultured for 4weeks at 37°C, and protein extraction was performed by sequential extraction. Bacterial cells were sonicated, centrifuged at 5000rpm for 45min, and the supernatant was collected and subjected to multiple rounds of treatment to prior to protein isolation...
December 2016: International Journal of Mycobacteriology
https://www.readbyqxmd.com/read/28042093/expression-and-antigenicity-of-recombinant-human-respiratory-syncytial-virus-glycoproteins-having-different-affinity-tags
#17
Han Saem Lee, A-Reum Kim, Kisoon Kim, Wan-Ji Lee, Sung Soon Kim, You-Jin Kim
Human respiratory syncytial virus (HRSV) is a main cause of lower respiratory tract infections in infants and the elderly. Glycoprotein (G) is major antigen on the viral surface, and plays a key role for virus entry. Therefore, purification of the glycoprotein of HRSV is critical for the development of HRSV vaccine and serological diagnosis. In this study, we report the design and characterization of glycoprotein engineered rationally to enhance the protein solubility and to facilitate efficient purification...
December 29, 2016: Protein Expression and Purification
https://www.readbyqxmd.com/read/28038927/cellular-immune-response-to-%C3%AE-2-glycoprotein-i-valine-leucine-247-phenotypes-in-mexican-patients-with-primary-antiphospholipid-syndrome
#18
Carlos A Núñez-Álvarez, Diego F Hernández-Ramírez, Araceli Martinez-Castillo, Virginia Pascual Ramos, Javier Cabiedes, Alicia Ortega, Antonio R Cabral
: Homozygote genotype V(247) of the β2-glycoprotein-I (β2GP-I) gene has been associated with anti-β2GP-I and thrombosis in patients with primary anti-phospholipid syndrome APS (PAPS). However, the cellular immune response to β2GP-I(247) has been little studied. OBJECTIVE: To evaluate the immune cellular proliferation in response to native and non-native β2GP-I(247) valine/leucine phenotype from Mexican patients with PAPS. METHODS: We studied 10 patients with PAPS and 10 healthy control subjects (HC)...
December 27, 2016: Human Immunology
https://www.readbyqxmd.com/read/28036315/evaluation-of-the-immunogenic-property-of-nt-h-influenzae-protein-d-with-neisseria-meningitidis-omv-in-balb-c
#19
Ava Behrouzi, Saeid Bouzari, Seyed Davar Siadat, Mana Oloomi, Mehdi Davari, Hoora Mazaheri
INTRODUCTION: Identifying ideal non typeable Haemophilus influenzae (NTHi) vaccine candidates has not been easy due to extensive sequence and antigenic variation among gene products interacting with the immune system. Protein D (PD) is a highly conserved 42 kDa surface lipoprotein available in all H. influenzae, including NTHi. METHODOLOGY: In this study, the gene encoding PD was cloned from H. influenzae and expressed in Escheriachia coli TOPO10 cell in pBAD vector...
December 30, 2016: Journal of Infection in Developing Countries
https://www.readbyqxmd.com/read/28033726/optimization-of-desferrioxamine-e-production-by-streptomyces-parvulus
#20
Tamás Gáll, Gábor Lehoczki, Gyöngyi Gyémánt, Tamás Emri, Zsuzsa M Szigeti, György Balla, József Balla, István Pócsi
Siderophores are produced by a number of microbes to capture iron with outstandingly high affinity, which property also generates biomedical and industrial interests. Desferrioxamine E (DFO-E) secreted by streptomycetes bacteria can be an ideal candidate for iron chelation therapy, which necessitates its cost-effective production for in vitro and animal studies. This study focused on the optimization of DFO-E production by Streptomyces parvulus CBS548.68. Different combinations of various carbon and nitrogen sources as well as the addition of 3-morpholinopropane-1-sulfonic acid (MOPS) markedly affected DFO-E yields, which were attributed, at least in part, to the higher biomass productions found in MOPS-supplemented cultures...
December 2016: Acta Microbiologica et Immunologica Hungarica
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