keyword
MENU ▼
Read by QxMD icon Read
search

purification

keyword
https://www.readbyqxmd.com/read/28641402/ec-tagging-allows-cell-type-specific-rna-analysis
#1
Naoki Hida, Mohamed Y Aboukilila, Dana A Burow, Rakesh Paul, Marc M Greenberg, Michael Fazio, Samantha Beasley, Robert C Spitale, Michael D Cleary
Purification of cell type-specific RNAs remains a significant challenge. One solution involves biosynthetic tagging of target RNAs. RNA tagging via incorporation of 4-thiouracil (TU) in cells expressing transgenic uracil phosphoribosyltransferase (UPRT), a method known as TU-tagging, has been used in multiple systems but can have limited specificity due to endogenous pathways of TU incorporation. Here, we describe an alternative method that requires the activity of two enzymes: cytosine deaminase (CD) and UPRT...
June 21, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28640955/in-solution-antibody-harvesting-with-a-plant-produced-hydrophobin-protein-a-fusion
#2
Katri Kurppa, Lauri Reuter, Anneli Ritala, Markus Linder, Jussi Joensuu
Purification is a bottleneck and a major cost factor in the production of antibodies. We set out to engineer a bi-functional fusion protein from two building blocks, Protein A and a hydrophobin, aiming at low-cost and scalable antibody capturing in solutions. Immunoglobulin-binding Protein A is widely used in affinity-based purification. The hydrophobin fusion tag, on the other hand, has been shown to enable purification by two-phase separation. Protein A was fused to two different hydrophobin tags, HFBI or II, and expressed transiently in Nicotiana benthamiana...
June 22, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28640945/production-of-human-vitronectin-in-nicotiana-benthamiana-using-the-inpact-hyper-expression-platform
#3
Benjamin Dugdale, Maiko Kato, Pradeep Deo, Manuel Plan, Mark Harrison, Robyn Lloyd, Terry Walsh, Robert Harding, James Dale
Human vitronectin (hVN) is a glycoprotein that functions as a cell adhesion molecule and a regulator of coagulation in blood plasma and the extracellular matrix. In vitro, hVN is added to serum-free media in order to promote the adhesion of animal cells to tissue culture surfaces and the proliferation of undifferentiated stem cells. Here, we report the production of hVN in Nicotiana benthamiana using the inducible In Plant Activation (INPACT) hyper-expression platform. N. benthamiana plants were transformed with an INPACT expression cassette encoding hVN, and both the Tobacco yellow dwarf virus Rep/RepA activator and Tomato bushy stunt virus p19 gene under the transcriptional control of the ethanol-inducible AlcR:alcA gene switch...
June 22, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28637286/structure-seq2-sensitive-and-accurate-genome-wide-profiling-of-rna-structure-in-vivo
#4
Laura E Ritchey, Zhao Su, Yin Tang, David C Tack, Sarah M Assmann, Philip C Bevilacqua
RNA serves many functions in biology such as splicing, temperature sensing, and innate immunity. These functions are often determined by the structure of RNA. There is thus a pressing need to understand RNA structure and how it changes during diverse biological processes both in vivo and genome-wide. Here, we present Structure-seq2, which provides nucleotide-resolution RNA structural information in vivo and genome-wide. This optimized version of our original Structure-seq method increases sensitivity by at least 4-fold and improves data quality by minimizing formation of a deleterious by-product, reducing ligation bias, and improving read coverage...
June 16, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28637209/purification-and-characterization-of-two-new-cell-bound-bioactive-compounds-produced-by-wildlactococcus-lactisstrain
#5
Margarete Alice Fontes Saraiva, Dag Anders Brede, Ingolf Figved Nes, Maria Cristina Baracat-Pereira, Marisa Vieira de Queiroz, Célia Alencar de Moraes
Novel compounds and innovative methods are required considering that antibiotic resistance has reached a crisis point. In the study, two cell-bound antimicrobial compounds produced by L. lactis ID1.5 were isolated and partially characterized. Following purification by cationic exchange and a solid phase C18 column, antimicrobial activity was recovered after three runs of RPC using 60% (v/v) and 100% (v/v) of 2-propanol for elution, suggesting that more than one antimicrobial compound were produced by L. lactis ID1...
June 20, 2017: FEMS Microbiology Letters
https://www.readbyqxmd.com/read/28636078/microbial-lipopeptide-production-and-purification-bioprocesses-current-progress-and-future-challenges
#6
REVIEW
François Coutte, Didier Lecouturier, Krasimir Dimitrov, Jean-Sébastien Guez, Frank Delvigne, Pascal Dhulster, Philippe Jacques
Lipopeoptides are amphiphilic compounds combining interesting physicochemical properties and biological activities. Due to their high foaming capacity in aerated bioreactor, the development of scalable bioprocesses for their production is a major bottleneck. In addition, the genes involved in the biosynthesis of these lipopeptides are mainly regulated by the quorum sensing, a global regulatory mechanism depending on cell density and known to be activated in biofilms. Several approaches have thus been considered in literature taking into account two criteria, on one side, to favor, control or avoid foam formation and on the other side, to use planktonic or immobilized (biofilm) cells...
June 21, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/28635655/from-farm-to-pharma-an-overview-of-industrial-heparin-manufacturing-methods
#7
REVIEW
Jan-Ytzen van der Meer, Edwin Kellenbach, Leendert J van den Bos
The purification of heparin from offal is an old industrial process for which commercial recipes date back to 1922. Although chemical, chemoenzymatic, and biotechnological alternatives for this production method have been published in the academic literature, animal-tissue is still the sole source for commercial heparin production in industry. Heparin purification methods are closely guarded industrial secrets which are not available to the general (scientific) public. However by reviewing the academic and patent literature, we aim to provide a comprehensive overview of the general methods used in industry for the extraction of heparin from animal tissue...
June 21, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/28635559/purification-of-antibody-against-ara-h-2-by-a-home-made-immunoaffinity-chromatography-column
#8
Zhihua Wu, Kun Li, Shaode Zhan, Ping Tong, Xin Li, Anshu Yang, Hongbing Chen
Antibodies are used extensively in numerous applications both in vivo and in vitro. To purify anti-Ara h 2 polyclonal antibody (PcaAb), a home-made immunoaffinity chromatography column (IAC) method was established. The properties of the home-made column were compared with those of the Mab Affinity Protein G Agarose High Flow (MPG), a commercially available column successfully used in capturing polyclonal antibodies. During antibody purification from rabbits antiserum against Ara h 2, the column capacity, recovery, and purification factor were characterized for IAC and MPG...
June 21, 2017: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/28635544/heparin-new-life-for-an-old-drug
#9
Carlos Raúl Aláez-Versón, Elena Lantero, Xavier Fernàndez-Busquets
Heparin is one of the oldest drugs, which nevertheless remains in widespread clinical use as an inhibitor of blood coagulation. The history of its identification a century ago unfolded amid one of the most fascinating scientific controversies turning around the distribution of credit for its discovery. The composition, purification and structure-function relationship of this naturally occurring glycosaminoglycan regarding its classical role as anticoagulant will be dealt with before proceeding to discuss its therapeutic potential in, among other, inflammatory and infectious disease, cancer treatment, cystic fibrosis and Alzheimer's disease...
June 21, 2017: Nanomedicine
https://www.readbyqxmd.com/read/28634775/hydrophilic-acylated-surface-protein-a-haspa-of-leishmania-donovani-expression-purification-and-biophysico-chemical-characterization
#10
Manoj Kumar, Kishu Ranjan, Vijay Singh, Chandramani Pathak, Anju Pappachan, Desh Deepak Singh
Hydrophilic acylated surface proteins (HASPs) are acidic surface proteins which get localized on the surface of Leishmania parasite during infective stages through a "non-classical" pathway. In this study, we report the heterologous expression and purification of Leishmania donovani HASPA (r-LdHASPA) in E. coli system and its partial characterization. The structural aspects of the purified protein were analyzed using CD spectroscopy and modeling studies which indicate that r-LdHASPA consists of random coils...
June 21, 2017: Protein Journal
https://www.readbyqxmd.com/read/28634757/a-uhplc-ms-ms-method-for-profiling-multifunctional-steroids-in-human-hair
#11
Zhen Dong, Caihong Wang, Jinlan Zhang, Zhe Wang
It is important to profile steroids in many physiological and pathological processes. Recently, hair has been used for the long-term measurement of endogenous steroid hormones. Analyzing hair has advantages of being noninvasive and time sequential compared with other bio-specimens. Liquid chromatography-mass spectrometry (LC-MS) techniques have been widely used over the past decades; however, it is challenging to profile estrogens in hair by LC-MS, and more comprehensive steroid profiling is required. In this paper, an ultra high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed to simultaneously profile 28 multifunctional steroids, including corticosteroids (n = 6), estrogens (n = 13), androgens (n = 5) and progestogens (n = 4), in human scalp hair in a single run...
June 20, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28634420/characterization-of-the-proinflammatory-profile-of-synovial-fluid-derived-exosomes-of-patients-with-osteoarthritis
#12
Rossana Domenis, Rossella Zanutel, Federica Caponnetto, Barbara Toffoletto, Adriana Cifù, Cinzia Pistis, Paolo Di Benedetto, Araldo Causero, Massimo Pozzi, Fabrizio Bassini, Martina Fabris, Kayvan R Niazi, Patrick Soon-Shiong, Francesco Curcio
The purpose of this study is to characterize synovial fluid- (SF-) derived exosomes of patients with gonarthrosis comparing two methods of isolation and to investigate their immune regulatory properties. Extracellular vesicles (EVs) have been isolated from inflamed SF by polymer precipitation method and quantified by Exocet kit and by nanoparticle tracking analysis. Vesicles expressed all the specific exosomal markers by immunoblot and FACS. After isolation with Exoquick, a relevant contamination by immune complexes was detected, which required further magnetic bead-based purification to remove...
2017: Mediators of Inflammation
https://www.readbyqxmd.com/read/28634114/peanut-digestome-identification-of-digestion-resistant-ige-binding-peptides
#13
Luigia Di Stasio, Gianluca Picariello, Mariantonietta Mongiello, Rita Nocerino, Roberto Berni Canani, Simona Bavaro, Linda Monaci, Pasquale Ferranti, Gianfranco Mamone
Stability to proteolytic degradation in the digestive tract is considered a general feature shared by most food allergens. Current digestibility models exclusively utilize purified allergen proteins, neglecting the relevant effects of matrix that occur for foodstuff systems. In the present study, we investigated digestion stability of the major peanut allergens directly in the natural matrix using an in vitro static model that simulates the gastrointestinal digestion including the oral, gastric, duodenal and intestinal (brush border membrane enzymes) phases...
June 17, 2017: Food and Chemical Toxicology
https://www.readbyqxmd.com/read/28633930/phospholipase-a2-in-the-venom-of-three-cottonmouth-snake-species
#14
Ying Jia, Boris Ermolinsky, Aryana Garza, Daniele Provenzano
Two phospholipase A2s (PLA2s), Asp49 (D49) and Lys49 (K49), were purified by one-step reverse-phase high performance liquid chromatography (RP-HPLC) from the venom of each of the three subspecies of cottonmouth snake, Western cottonmouth (Agkistrodon piscivorus leucostoma; Apl), Eastern cottonmouth (Agkistrodon piscivorus piscivorus; App) and Florida cottonmouth (Agkistrodon piscivorus conanti; Apc). Venom protein profiles and PLA2s elution pattern of the three cottonmouth snakes were remarkably similar displaying four similar sharp and two wide peaks; in all cases K49 PLA2 eluted first followed by D49 PLA2...
June 17, 2017: Toxicon: Official Journal of the International Society on Toxinology
https://www.readbyqxmd.com/read/28633820/green-synthesis-of-ag-cr-ac-nanocomposites-by-azadirachta-indica-and-its-application-for-the-simultaneous-removal-of-binary-mixture-of-dyes-by-ultrasonicated-assisted-adsorption-process-using-response-surface-methodology
#15
Muhammad Saad, Hajira Tahir, Duaa Ali
In the present studies the Ag-Cr-AC nanocomposites were synthesized by Azadirachta indica leaves extract. They were inoculated on the amorphous surface of activated carbon. The surface morphology and structural identification was determined by SEM, FTIR and XRD techniques. The simultaneous removal of binary dye system of Reactive Red and Crystal Violet were performed by ultrasonicated assisted adsorption process utilizing Ag-Cr-AC nanocomposites. Central Composite Design (CCD) having 5 factors of time, pH, amount of Ag-Cr-AC (adsorbent), concentrations of Reactive Red (RR) and Crystal Violet (CV) was employed...
September 2017: Ultrasonics Sonochemistry
https://www.readbyqxmd.com/read/28632821/encapsulated-islet-transplantation-where-do-we-stand
#16
Vijayaganapathy Vaithilingam, Sumeet Bal, Bernard E Tuch
Transplantation of pancreatic islets encapsulated within immuno-protective microcapsules is a strategy that has the potential to overcome graft rejection without the need for toxic immunosuppressive medication. However, despite promising preclinical studies, clinical trials using encapsulated islets have lacked long-term efficacy, and although generally considered clinically safe, have not been encouraging overall. One of the major factors limiting the long-term function of encapsulated islets is the host's immunological reaction to the transplanted graft which is often manifested as pericapsular fibrotic overgrowth (PFO)...
2017: Review of Diabetic Studies: RDS
https://www.readbyqxmd.com/read/28631997/expression-and-purification-of-a-functional-porcine-soluble-triggering-receptor-expressed-on-myeloid-cells-1
#17
Hui Li, Shuangshuang Guo, Leyan Yan, Chunhua Meng, Yiyi Hu, Kongwang He, Zhendan Shi
Triggering receptor expressed on myeloid cells 1 (TREM-1) plays a vital role in the pathogen-triggered amplification loop required for proinflammatory responses. Blockade of TREM-1 signaling may inhibit expansion of sepsis and prolong survival of animals. In the present study, the gene of porcine soluble TREM-1 was cloned and expressed in E. coli. After purification, the bioactivity of recombinant porcine soluble TREM-1 was tested in vitro on porcine alveolar macrophages. The results showed that supplementation with the recombinant porcine sTREM-1 protein rapidly and dose-dependently attenuated the upregulation of cytokines (IL-1β, IL-2, IL-4, IL-8, IL-10, IL-12, IL-16, IL-18, and TNF-α) caused by LPS stimulation in the cultured porcine alveolar macrophages...
February 10, 2017: Animal Biotechnology
https://www.readbyqxmd.com/read/28631792/nanoscopic-dynamics-of-bicontinous-microemulsions-effect-of-membrane-associated-protein
#18
V K Sharma, Douglas G Hayes, Volker S Urban, Hugh M O'Neill, M Tyagi, E Mamontov
Bicontinous microemulsions (BμE) generally consist of nanodomains formed by surfactant in a mixture of water and oil at nearly equal proportions and are potential candidates for the solubilization and purification of membrane proteins. Here we present the first time report of nanoscopic dynamics of surfactant monolayers within BμEs formed by the anionic surfactant sodium dodecyl sulfate (SDS) measured on the nanosecond to picosecond time scale using quasielastic neutron scattering (QENS). BμEs investigated herein consisted of middle phases isolated from Winsor-III microemulsion systems that were formed by mixing aqueous and oil solutions under optimal conditions...
June 20, 2017: Soft Matter
https://www.readbyqxmd.com/read/28631591/comparative-evaluation-of-veriflow-%C3%A2-listeria-species-to-usda-culture-based-method-for-the-detection-of-listeria-spp-in-food-and-environmental-samples
#19
Adam C Joelsson, Shawn P Terkhorn, Ashley S Brown, Amrita Puri, Benjamin J Pascal, Zara E Gaudioso, Nicholas A Siciliano
Veriflow(®)Listeria species (Veriflow LS) isa molecular-based assay for the presumptive detection of Listeria spp. from environmental surfaces (stainless steel, sealed concrete, plastic, and ceramic tile) and ready-to-eat (RTE) food matrixes (hot dogs and deli meat). The assay utilizes a PCR detection method coupled with a rapid, visual, flow-based assay that develops in 3 min post-PCR amplification and requires only a 24 h enrichment for maximum sensitivity. The Veriflow LS system eliminates the need for sample purification, gel electrophoresis, or fluorophore-based detection of target amplification and does not require complex data analysis...
June 20, 2017: Journal of AOAC International
https://www.readbyqxmd.com/read/28631590/comparative-evaluation-of-veriflow-%C3%A2-salmonella-species-to-usda-and-fda-culture-based-methods-for-the-detection-of-salmonella-spp-in-food-and-environmental-samples
#20
Amrita Puri, Adam C Joelsson, Shawn P Terkhorn, Ashley S Brown, Zara E Gaudioso, Nicholas A Siciliano
Veriflow(®)Salmonella species (Veriflow SS)is a molecular-based assay for the presumptive detection of Salmonella spp. from environmental surfaces (stainless steel, sealed concrete, plastic, and ceramic tile), dairy (2% milk), raw meat (20% fat ground beef), chicken carcasses, and ready-to-eat (RTE) food (hot dogs). The assay utilizes a PCR detection method coupled with a rapid, visual, flow-based assay that develops in 3 min post-PCR amplification and requires only an 18 h enrichment for maximum sensitivity...
June 20, 2017: Journal of AOAC International
keyword
keyword
28154
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"