keyword
MENU ▼
Read by QxMD icon Read
search

protein purification

keyword
https://www.readbyqxmd.com/read/28346045/a-modular-and-adaptive-mass-spectrometry-based-platform-for-support-of-bioprocess-development-toward-optimal-host-cell-protein-clearance
#1
Donald E Walker, Feng Yang, Joseph Carver, Koman Joe, David A Michels, X Christopher Yu
A modular and adaptive mass spectrometry (MS)-based platform was developed to provide fast, robust and sensitive host cell protein (HCP) analytics to support process development. This platform relies on one-dimensional ultra-high performance liquid chromatography (1D UHPLC) combined with several different MS data acquisition strategies to meet the needs of purification process development. The workflow was designed to allow HCP composition and quantitation for up to 20 samples per day, a throughput considered essential for real time bioprocess development support...
March 27, 2017: MAbs
https://www.readbyqxmd.com/read/28345863/native-mass-spectrometry-of-recombinant-proteins-from-crude-cell-lysates
#2
Jinrui Gan, Gili Ben-Nissan, Galina Arkind, Mark Tarnavsky, Devin Trudeau, Lianet Noda Garcia, Dan Tawfik, Michal Sharon
Determining the properties of proteins prior to purification saves time and labor. Here, we demonstrate a native mass spectrometry approach for rapid characterization of overexpressed proteins directly in crude cell lysates. The method provides immediate information on the identity, solubility, oligomeric state, overall structure and stability, as well as ligand binding, without the need for purification.
March 27, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28345298/effects-of-t-loop-modification-on-the-pii-signaling-protein-structure-of-uridylylated-escherichia-coli-glnb-bound-to-atp
#3
Carles Palanca, Vicente Rubio
To adapt to environments with variable nitrogen sources and richness, the widely distributed homotrimeric PII signaling proteins bind their allosteric effectors ADP/ATP/2-oxoglutarate, and experience nitrogen-sensitive uridylylation of their flexible T-loops at Tyr51, regulating their interactions with effector proteins. To clarify whether uridylylation triggers a given T-loop conformation, we determined the crystal structure of the classical paradigm of PII protein, Escherichia coli GlnB (EcGlnB), in fully uridylylated form (EcGlnB-UMP3 )...
March 26, 2017: Environmental Microbiology Reports
https://www.readbyqxmd.com/read/28343996/optimization-of-overexpression-of-a-chaperone-protein-of-steroid-c25-dehydrogenase-for-biochemical-and-biophysical-characterization
#4
Ewa Niedzialkowska, Beata Mrugała, Agnieszka Rugor, Mateusz P Czub, Anna Skotnicka, Julien J H Cotelesage, Graham N George, Maciej Szaleniec, Wladek Minor, Krzysztof Lewiński
Molybdenum is an essential nutrient for plant, bacteria and animal metabolism. Molybdoenzymes are involved in nitrogen assimilation and oxidoreductive, detoxification and bioconversion reactions of environmental, industrial, and pharmaceutical interest. Molybdoenzymes contain a molybdenum cofactor (Moco) which is a pyranopterin heterocyclic compound that binds a molybdenum atom via a dithiolene group. Because Moco is a large and complex compound deeply buried within protein, molybdoenzymes are accompanied by their private chaperone protein responsible for the cofactor's insertion into the enzyme and the enzyme's maturation...
March 23, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28343687/multiple-simultaneous-independent-gradients-for-a-versatile-multidimensional-liquid-chromatography-part-ii-application-2-computer-controlled-ph-gradients-in-the-presence-of-urea-provide-improved-separation-of-proteins-stability-influenced-anion-and-cation-exchange
#5
Allen G Hirsh, Latchezar I Tsonev
This paper details the use of a method of creating controlled pH gradients (pISep) to improve the separation of protein isoforms on ion exchange (IEX) stationary phases in the presence of various isocratic levels of urea. The pISep technology enables the development of computer controlled pH gradients on both cationic (CEX) and anionic (AEX) IEX stationary phases over the very wide pH range from 2 to 12. In pISep, titration curves generated by proportional mixing of the acidic and basic pISep working buffers alone, or in the presence of non-buffering solutes such as the neutral salt NaCl (0-1M), polar organics such as urea (0-8M) or acetonitrile (0-80 Vol%), can be fitted with high fidelity using high order polynomials which, in turn allows construction of a mathematical manifold %A (% acidic pISep buffer) vs...
March 16, 2017: Journal of Chromatography. A
https://www.readbyqxmd.com/read/28342968/heterologous-expression-purification-and-characterization-of-three-novel-esterases-secreted-by-the-lignocellulolytic-fungus-penicillium-purpurogenum-when-grown-on-sugar-beet-pulp
#6
Gabriela Oleas, Eduardo Callegari, Romina Sepúlveda, Jaime Eyzaguirre
The lignocellulolytic fungus, Penicillium purpurogenum, grows on a variety of natural carbon sources, among them sugar beet pulp. Culture supernatants of P. purpurogenum grown on sugar beet pulp were partially purified and the fractions obtained analyzed for esterase activity by zymograms. The bands with activity on methyl umbelliferyl acetate were subjected to mass spectrometry to identify peptides. The peptides obtained were probed against the proteins deduced from the genome sequence of P. purpurogenum...
March 18, 2017: Carbohydrate Research
https://www.readbyqxmd.com/read/28342912/peanut-oleosins-associated-with-severe-peanut-allergy-importance-of-lipophilic-allergens-for-comprehensive-allergy-diagnostics
#7
Christian Schwager, Skadi Kull, Jochen Behrends, Niels Röckendorf, Frauke Schocker, Andreas Frey, Arne Homann, Wolf-Meinhard Becker, Uta Jappe
BACKGROUND: Peanut allergy is one of the most common and most severe food allergies in Western countries and its accurate diagnosis to prevent potential life-threatening allergic reactions is crucial. However, aqueous extracts used for routine diagnostic measurements are devoid of lipophilic allergens such as oleosins. We have recently succeeded in the isolation and purification of these unique proteins, and the current study evaluates their allergenic potential and clinical relevance...
March 22, 2017: Journal of Allergy and Clinical Immunology
https://www.readbyqxmd.com/read/28342667/a-fast-and-efficient-purification-platform-for-cell-based-influenza-viruses-by-flow-through-chromatography
#8
Yu-Fen Tseng, Tsai-Chuan Weng, Chia-Chun Lai, Po-Ling Chen, Min-Shi Lee, Alan Yung-Chih Hu
Since newly emerging influenza viruses with pandemic potentials occurred in recent years, the demand for producing pandemic influenza vaccines for human use is high. For the development of a quick and efficient vaccine production, we proposed an efficient purification platform from the harvest to the purified bulk for the cell-based influenza vaccine production. This platform based on flow-through chromatography and filtration steps and the process only involves a few purification steps, including depth filtration, inactivation by formaldehyde, microfiltration, ultrafiltration, anion-exchange and ligand-core chromatography and sterile filtration...
March 22, 2017: Vaccine
https://www.readbyqxmd.com/read/28342173/metal-ions-binding-t4-lysozyme-as-an-intramolecular-protein-purification-tag-compatible-with-x-ray-crystallography
#9
Evzen Boura, Adriana Baumlova, Dominika Chalupska, Anna Dubankova, Martin Klima
Phage T4 lysozyme is a well folded and highly soluble protein that is widely used as an insertion tag to improve solubility and crystallization properties of poorly behaved recombinant proteins. It has been used in the fusion protein strategy to facilitate crystallization of various proteins including multiple G protein-coupled receptors, lipid kinases, or sterol binding proteins. Here, we present a structural and biochemical characterization of its novel, metal ions-binding mutant (mbT4L). We demonstrate that mbT4L can be used as a purification tag in the immobilized-metal affinity chromatography and that, in many respects, it is superior to the conventional hexahistidine tag...
March 25, 2017: Protein Science: a Publication of the Protein Society
https://www.readbyqxmd.com/read/28340724/mass-spectrometry-and-metallomics-a-general-protocol-to-assess-stability-of-metallodrug-protein-adducts-in-bottom-up-ms-experiments
#10
Elena Michelucci, Giuseppe Pieraccini, Gloriano Moneti, Chiara Gabbiani, Alessandro Pratesi, Luigi Messori
The bottom-up mass spectrometry approach is today one of the best tools of Metallomics to characterize the binding of metal-based drugs to proteins. Yet, the stability of metal-protein coordination bonds along the whole process may be a critical issue. This led us to build up a general protocol to test metallodrug-protein adduct stability under the typical conditions of the filter-aided sample preparation (FASP)/bottom-up procedure, ranging from the analysis of solutions containing metal-protein adducts to tandem mass spectrometry experiments...
May 15, 2017: Talanta
https://www.readbyqxmd.com/read/28340620/stable-high-level-expression-of-factor-viii-in-chinese-hamster-ovary-cells-in-improved-elongation-factor-1-alpha-based-system
#11
Nadezhda A Orlova, Sergey V Kovnir, Alexandre G Gabibov, Ivan I Vorobiev
BACKGROUND: Recombinant factor VIII (FVIII), used for haemophilia A therapy, is one of the most challenging among the therapeutic proteins produced in heterologous expression systems. Deletion variant of FVIII, in which the entire domain B is replaced by a short linker peptide, was approved for medical use. Efficacy and safety of this FVIII deletion variant are similar to full-length FVIII preparations while the level of production in CHO cells is substantially higher. Typical levels of productivity for CHO cell lines producing deletion variant FVIII-BDD SQ, described elsewhere, are 0...
March 24, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28339303/development-of-rap-tag-a-novel-tagging-system-for-protein-detection-and-purification
#12
Yuki Fujii, Mika K Kaneko, Satoshi Ogasawara, Shinji Yamada, Miyuki Yanaka, Takuro Nakamura, Noriko Saidoh, Kanae Yoshida, Ryusuke Honma, Yukinari Kato
Affinity tag systems, possessing high affinity and specificity, are useful for protein detection and purification. The most suitable tag for a particular purpose should be selected from many available affinity tag systems. In this study, we developed a novel affinity tag called the "RAP tag" system, which comprises a mouse antirat podoplanin monoclonal antibody (clone PMab-2) and the RAP tag (DMVNPGLEDRIE). This system is useful not only for protein detection in Western blotting, flow cytometry, and sandwich enzyme-linked immunosorbent assay, but also for protein purification...
March 24, 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28338731/purification-and-enzymatic-characterization-of-gallus-gallus-blm-helicase
#13
Jing Shi, Na-Nv Liu, Yan-Tao Yang, Xu-Guang Xi
Mutations in human BLM helicase give rise to the autosomal recessive Bloom syndrome, which shows high predisposition to types of malignant tumours. Though lots of biochemical and structural investigations have shed lights on the helicase core, structural investigations of the whole BLM protein are still limited due to its low stability and production. Here by comparing with the expression systems and functions of other BLM homologues, we developed the heterologous high-level expression and high-yield purification systems for Gallus gallus BLM (gBLM) in Escherichia coli...
February 21, 2017: Journal of Biochemistry
https://www.readbyqxmd.com/read/28338200/glioma-cells-promote-angiogenesis-through-the-release-of-exosomes-containing-long-non-coding-rna-pou3f3
#14
H-L Lang, G-W Hu, Y Chen, Y Liu, W Tu, Y-M Lu, L Wu, G-H Xu
OBJECTIVE: Angiogenesis is a key event in the progression of gliomas, and emerging evidence suggests that exosomes are signaling extracellular organelles that modulate the tumor microenvironment and promote angiogenesis and tumor progression. This study aimed to explore the mechanism by which glioma-derived exosomes affect angiogenesis. MATERIALS AND METHODS: qRT-PCR was used to determine the expression level of linc-POU3F3 in glioma tissue as well as glioma cell lines...
March 2017: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/28337691/purification-and-characterization-of-elizabethkingia-l-amino-acid-esterase-an-enzyme-useful-for-enzymatic-synthesis-of-the-dipeptide-valyl-glycine
#15
Takahiro Tanaka, Kazuyoshi Takagi, Hossain Md Saddam, Yoichi Takeda, Mamoru Wakayama
Valyl-glycine (Val-Gly) is useful as a synthetic substrate of γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly), which exhibits a strong taste of "kokumi." For efficient enzymatic synthesis of Val-Gly from valine methylester and glycine using L-amino acid esterase (LAE), we screened microorganisms producing LAE with synthetic activity toward Val-Gly. Among 17 isolates showing LAE activity, Elizabethkingia sp. TT1, which was identified by 16S rDNA sequence analysis, showed the highest synthetic activity toward Val-Gly...
March 24, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28334977/sclip-an-integrated-platform-to-study-rna-protein-interactomes-in-biomedical-research-identification-of-cstf2tau-in-alternative-processing-of-small-nuclear-rnas
#16
Yulia Kargapolova, Michal Levin, Karl Lackner, Sven Danckwardt
RNA-binding proteins (RBPs) are central for gene expression by controlling the RNA fate from birth to decay. Various disorders arising from perturbations of RNA-protein interactions document their critical function. However, deciphering their function is complex, limiting the general functional elucidation of this growing class of proteins and their contribution to (patho)physiology. Here, we present sCLIP, a simplified and robust platform for genome-wide interrogation of RNA-protein interactomes based on crosslinking-immunoprecipitation and high-throughput sequencing...
February 28, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28330749/identification-purification-and-expression-patterns-of-chitinase-from-psychrotolerant-pedobacter-sp-pr-m6-and-antifungal-activity-in%C3%A2-vitro
#17
Yong-Su Song, Dong-Jun Seo, Woo-Jin Jung
In this study, a novel psychrotolerant chitinolytic bacterium Pedobacter sp. PR-M6 that displayed strong chitinolytic activity on 0.5% colloidal chitin was isolated from the soil of a decayed mushroom. Chitinase activity of PR-M6 at 25 °C (C25) after 6 days of incubation with colloidal chitin increased rapidly to a maximum level (31.3 U/mg proteins). Three chitinase isozymes (chiII, chiIII, and chiIV) from the crude enzyme at 25 °C (C25) incubation were expressed on SDS-PAGE gels at 25 °C. After purification by chitin-affinity chromatography, six chitinase isozymes (chiI, chiII, chiIII, chiIV, chiV, and chiVI) from C25-fractions were expressed on SDS-PAGE gels at 25 °C...
March 19, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/28330285/screening-production-optimization-and-characterization-of-%C3%AE-glucosidase-using-microbes-from-shellfish-waste
#18
Samhita Mahapatra, A S Vickram, T B Sridharan, R Parameswari, M Ramesh Pathy
An extracellular β-glucosidase was isolated from Proteus mirabilis VIT117 found to be growing on prawn shells. The enzyme production was found to be enhanced (14.58 U/ml) when the culture was maintained at pH 9 and provided with sorbitol as carbon source, yeast extract as nitrogen source and incubated at 37 °C for approximately 72 h. Statistical methods like Plackett-Burman and RSM were also applied here to study the effects of different combinations of growth parameters for the bacteria, where the most significant parameters were found to be inoculum size, pH, yeast extract, incubation time and sorbitol...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28330275/phytochemicals-of-moringa-oleifera-a-review-of-their-nutritional-therapeutic-and-industrial-significance
#19
REVIEW
Ramesh Kumar Saini, Iyyakkannu Sivanesan, Young-Soo Keum
Moringa oleifera Lam., also known as the 'drumstick tree,' is recognized as a vibrant and affordable source of phytochemicals, having potential applications in medicines, functional food preparations, water purification, and biodiesel production. The multiple biological activities including antiproliferation, hepatoprotective, anti-inflammatory, antinociceptive, antiatherosclerotic, oxidative DNA damage protective, antiperoxidative, cardioprotective, as well as folk medicinal uses of M. oleifera (MO) are attributed to the presence of functional bioactive compounds, such as phenolic acids, flavonoids, alkaloids, phytosterols, natural sugars, vitamins, minerals, and organic acids...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28330270/expression-and-purification-of-a-gene-encoding-a-9-7%C3%A2-kda-pe-protein-of-mycobacterium-avium-subsp-paratuberculosis
#20
S Chandra Sekar, P P Goswami, R Deb
Mycobacterium avium subsp. paratuberculosis (Map) contains PE family antigens which are Proline and glutamic acid rich and may play important role as T-cell antigens. In the present study, the Map 1507 ORF encoding 9.7 kDa PE protein was amplified by polymerase chain reaction and cloned into E. coli vector pQE30 UA. The recombinant plasmid designated as pQ PE was transformed into E. coli M15 cells and induced with IPTG revealed the high level expression of 11.9 kDa His-fusion protein as estimated by migration in 15 % sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)...
December 2016: 3 Biotech
keyword
keyword
32512
1
2
Fetch more papers »
Fetching more papers... Fetching...
Read by QxMD. Sign in or create an account to discover new knowledge that matter to you.
Remove bar
Read by QxMD icon Read
×

Search Tips

Use Boolean operators: AND/OR

diabetic AND foot
diabetes OR diabetic

Exclude a word using the 'minus' sign

Virchow -triad

Use Parentheses

water AND (cup OR glass)

Add an asterisk (*) at end of a word to include word stems

Neuro* will search for Neurology, Neuroscientist, Neurological, and so on

Use quotes to search for an exact phrase

"primary prevention of cancer"
(heart or cardiac or cardio*) AND arrest -"American Heart Association"