scholarly journals Amplification and Purification of T4-Like Escherichia coli Phages for Phage Therapy: from Laboratory to Pilot Scale

2013 ◽  
Vol 80 (4) ◽  
pp. 1469-1476 ◽  
Author(s):  
Gilles Bourdin ◽  
Bertrand Schmitt ◽  
Laure Marvin Guy ◽  
Jacques-Edouard Germond ◽  
Sophie Zuber ◽  
...  
Keyword(s):  
High Speed ◽  
Production Systems ◽  
Anion Exchange Chromatography ◽  
Pilot Scale ◽  
Exchange Chromatography ◽  
Cold Chain ◽  
Contamination Level ◽  
Stirred Tank Reactors ◽  
Medium Speed ◽  
Strong Anion Exchange

ABSTRACTWe investigated the amplification and purification of phage preparations with respect to titer, contamination level, stability, and technical affordability. Using various production systems (wave bags, stirred-tank reactors, and Erlenmeyer flasks), we obtained peak titers of 109to 1010PFU/ml for T4-like coliphages. Phage lysates could be sterilized through 0.22-μm membrane filters without titer loss. Phages concentrated by differential centrifugation were not contaminated with cellular debris or bacterial proteins, as assessed by electron microscopy and mass spectrometry, respectively. Titer losses occurred by high-speed pelleting of phages but could be decreased by sedimentation through a sucrose cushion. Alternative phage concentration methods are prolonged medium-speed centrifugation, strong anion-exchange chromatography, and ultrafiltration, but the latter still allowed elevated lipopolysaccharide contamination. T4-like phages could not be pasteurized but maintained their infectivity titer in the cold chain. In the presence of 10 mM magnesium ions, phages showed no loss of titer over 1 month at 30°C.

scholarly journals Specific Non-Reducing Ends in Heparins from Different Animal Origins: Building Blocks Analysis Using Reductive Amination Tagging by Sulfanilic Acid

Molecules ◽  
2020 ◽  
Vol 25 (23) ◽  
pp. 5553
Author(s):  
Pierre A. J. Mourier
Keyword(s):  
Building Block ◽  
Reductive Amination ◽  
Building Blocks ◽  
Anion Exchange Chromatography ◽  
Sulfanilic Acid ◽  
Exchange Chromatography ◽  
Sulfated Polysaccharides ◽  
Liquid Chromatography Mass Spectrometry ◽  
Anticoagulant Drugs ◽  
Strong Anion Exchange

Heparins are linear sulfated polysaccharides widely used as anticoagulant drugs. Their nonreducing-end (NRE) has been little investigated due to challenges in their characterization, but is known to be partly generated by enzymatic cleavage with heparanases, resulting in N-sulfated glucosamines at the NRE. Uronic NRE (specifically glucuronic acids) have been isolated from porcine heparin, with GlcA-GlcNS,3S,6S identified as a porcine-specific NRE marker. To further characterize NRE in heparinoids, a building block analysis involving exhaustive heparinase digestion and subsequent reductive amination with sulfanilic acid was performed. This study describes a new method for identifying heparin classical building blocks and novel NRE building blocks using strong anion exchange chromatography on AS11 columns for the assay, and ion-pair liquid chromatography-mass spectrometry for building block identification. Porcine, ovine, and bovine intestine heparins were analyzed. Generally, NRE on these three heparins are highly sulfated moieties, particularly with 3-O sulfates, and the observed composition of the NRE is highly dependent on heparin origin. At the highest level of specificity, the isolated marker was only detected in porcine heparin. However, the proportion of glucosamines in the NRE and the proportion of glucuronic/iduronic configurations in the NRE uronic moieties greatly varied between heparin types.

scholarly journals Heparin Isomeric Oligosaccharide Separation Using Volatile Salt Strong Anion Exchange Chromatography

2016 ◽  
Vol 88 (23) ◽  
pp. 11542-11550 ◽  
Author(s):  
Rebecca L. Miller ◽  
Scott E. Guimond ◽  
Maitreyi Shivkumar ◽  
Jemma Blocksidge ◽  
James A. Austin ◽  
...  
Keyword(s):  
Anion Exchange ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Strong Anion Exchange

scholarly journals Comparison of Sugar Profile between Leaves and Fruits of Blueberry and Strawberry Cultivars Grown in Organic and Integrated Production System

Plants ◽  
2019 ◽  
Vol 8 (7) ◽  
pp. 205 ◽  
Author(s):  
Milica Fotirić Akšić ◽  
Tomislav Tosti ◽  
Milica Sredojević ◽  
Jasminka Milivojević ◽  
Mekjell Meland ◽  
...  
Keyword(s):  
High Performance ◽  
Production Systems ◽  
Amperometric Detection ◽  
Principal Component ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Defense Strategies ◽  
Integrated Production ◽  
Profile Distribution ◽  
Sugar Profile

The objective of this study was to determine and compare the sugar profile, distribution in fruits and leaves and sink-source relationship in three strawberry (‘Favette’, ‘Alba’ and ‘Clery’) and three blueberry cultivars (‘Bluecrop’, ‘Duke’ and ‘Nui’) grown in organic (OP) and integrated production systems (IP). Sugar analysis was done using high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection (PAD). The results showed that monosaccharide glucose and fructose and disaccharide sucrose were the most important sugars in strawberry, while monosaccharide glucose, fructose, and galactose were the most important in blueberry. Source-sink relationship was different in strawberry compared to blueberry, having a much higher quantity of sugars in its fruits in relation to leaves. According to principal component analysis (PCA), galactose, arabinose, and melibiose were the most important sugars in separating the fruits of strawberries from blueberries, while panose, ribose, stachyose, galactose, maltose, rhamnose, and raffinose were the most important sugar component in leaves recognition. Galactitol, melibiose, and gentiobiose were the key sugars that split out strawberry fruits and leaves, while galactose, maltotriose, raffinose, fructose, and glucose divided blueberry fruits and leaves in two groups. PCA was difficult to distinguish between OP and IP, because the stress-specific responses of the studied plants were highly variable due to the different sensitivity levels and defense strategies of each cultivar, which directly affected the sugar distribution. Due to its high content of sugars, especially fructose, the strawberry cultivar ‘Clery’ and the blueberry cultivars ‘Bluecrop’ and ‘Nui’ could be singled out in this study as being the most suitable cultivars for OP.

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