Chromatography polysaccharides

Affinity chromatography is used in the preparation of more highly purified Factor IX concentrates (53—55) as well as in the preparation of products such as antithrombin III [9000-94-6] (56,57). Heparin [9005-49-6], a sulfated polysaccharide (58), is the ligand used most commonly in these appHcations because it possesses specific binding sites for a number of plasma proteins (59,60). 

The separation of the polysaccharide components utilizes their different solubUities, polar groups, extents of branching, molecular weights, and molecular flexibUities and may be accompUshed batchwise or with easUy automated column techniques such as column or high performance Uquid chromatography. These procedures have been summarized in several reviews (3,141—143). 

Analytical techniques that utilise biopolymers, ie, natural macromolecules such as proteias, nucleic acids, and polysaccharides that compose living substances, represent a rapidly expanding field. The number of appHcations is large and thus uses hereia are limited to chiral chromatography, immunology, and biosensors. 

Lipoteichoic acids (from gram-positive bacteria) [56411-57-5J. Extracted by hot phenol/water from disrupted cells. Nucleic acids that were also extracted were removed by treatment with nucleases. Nucleic resistant acids, proteins, polysaccharides and teichoic acids were separated from lipoteichoic acids by anion-exchange chromatography on DEAE-Sephacel or by hydrophobic interaction on octyl-Sepharose [Fischer et al. Ear J Biochem 133 523 1983]. 

Nonionic polysaccharides are one of the most simple substances to analyze by size exclusion chromatography because they seldom exhibit nonsize exclusion effects. Due to their wide molecular weight distribution, TSK-GEL PW columns are recommended for their analysis. 

ANALYTICAL AND PREPARATIVE COLUMNS FOR AQUEOUS SIZE EXCLUSION CHROMATOGRAPHY OF POLYSACCHARIDES... 

J. Dingenen, Polysaccharide phases in enantioseparations in A practical approach to chiral separations by liquid chromatography, G. Subramanian, VCH, Weinheim (1994) Chapter 6. 

Shortly afterwards, Westphal, Liideritz, and their coworkers using the newly developed method of paper chromatography, found a new class of sugars in lipopolysaccharides (LPS) from Gram-negative bacteria, and identified them as 3,6-dideoxyhexoses. This work is summarized in Ref. 4. These discoveries initiated more-systematic investigations of hydrolyzates from bacterial polysaccharides, and a number of new monosaccharides were completely or partially identified. This development has been summarized by Ashwell and Hickman. ... 

Total polysaccharides were recovered in the ultrafiltration retentates on a Carbosep M5 membrane (Tech-Sep, MWCO 20 kDa) in the case of the red wine, or on a Centricon 30 membrane (Amicon, MWCO 30 kDa) in the ease of the apple and tomato juices. RG-II purification from the total polysaccharide coneentrates included, if necessary, several chromatography steps ... 

Further research is directed to the determination of the fine-structure of the pectins and hemicelluloses isolated from soy meal, using chromatography and degradation with specific enzymes. With these results a model of the polysaccharides present in the cell wall of soy will be formulated. Furthermore, application directed experiments will be performed to obtain information about structure-function relationships. 

The leaves of large plantain Plantago major L.) are used for wotmd healing in the traditional medicine. The effect might be due to biologically active polysaccharides. A pectin, PMII with anti-complementary activity has been isolated from the leaves by water extraction and ion exchange chromatography (1). 

Gel chromatography on Sephadex GlOO (2.8x50cm) of polysaccharide fraction I (Sample 2). The polysaccharide fraction was dissolved in a phosphate buffer at pH 6. After centrifugation, the supernatant was applied to the column at a flow rate of 0.8 ml/min. The elution was performed with a phosphate buffer and fractions of 10 ml each were collected. The refraction of each fraction was measured interferometrically. Fractions with coincident peaks were collected and analyzed for content of galacrturonic acid, neutral sugars and protein. 

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