Electrophoresis of polysaccharides

Zone electrophoresis is mostly used for biological applications. Peptide separation and the measurement of protein fractions from blood serum (proteinogram of albumin and o-, (3- and 7-globulins) are among the better known applications. This TLC for biochemists is useful for the separation of polysaccharides, nucleic acids (for DNA sequencing), proteins and other colloidal species. 

The development of electrophoretic techniques afforded possibilities for fractionations based on charge density differences. Duxbury (1989) has reviewed applications of different electrophoretic separation methods, including zone electrophoresis, moving boundary electrophoresis, isotachophoresis, and isoelectric focusing (IEF). Preparative column electrophoresis (Clapp, 1957) and continuous flow paper electrophoresis (Hayes, 1960 summarized by Hayes et al., 1985) methods have been used to separate components isolated from sapric histosol soils. These techniques allowed separation of polysaccharides from the colored components the electrophoretograms of the colored components were diffuse, showing a continuum of components of different charge densities. 

Jann, B., Reske, K., Jann, K. Heterogeneity of lipopolysaccharides. Analysis of polysaccharide chain lengths by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Eur J Biochem 60 (1975) 239-246. 

Chromatography of polysaccharides on silica gel and on carbon columns has been used to a limited extent. The use of ion-exchange resins has been investigated by Deuel and coworkers. The colunms had low capacities, and part of the material appeared to be irreversibly adsorbed. Deuel and coworkers, however, have reported better results when columns of cationic derivatives of cellulose, for instance (diethylaminoethyl)-cellulose, are used. Electrophoresis both by the Tiselius method on columns and on filter paper or glass-fiber sheets can give good separa-... 

Electron microscopy, studies on dex-trans, 349 Electrophoresis, 29 of polysaccharides, 278 Erythritol, from polysaccharides, 87 Erythrose, D-, 119, 251 osazone, 81... 

The shape of the precipitation curve obtained on gradual addition of ethanol to an aqueous solution of the polysaccharide material indicated heterogeneity. However, the five fractions collected showed no significant quantitative differences in component sugars. Another attempt to obtain fractionation of the polysaccharides by precipitation with ethanol from a solution in water or formamide was unsuccessful. Precipitation with hexadecyltrimethylammonium bromide likewise produced no clear separation. 2 Electrophoresis of some preparations showed them to be... 

These results emphasize the value of results obtained by the application of zone electrophoresis in the comparison of polysaccharides of supposedly similar chemical structure. 

Antibodies to polysaccharide antigens such as dextrans and levans produced in humans, while heterogeneous, often show restricted heterogeneity as shown by acrylamide gel electrophoresis (Yount et al., 1968), isoelectric focusing patterns (Cisar et al., 1975), starch gel electrophoresis of the separated polypeptide chains (Edelman and Kabat, 1964) and in possessing fewer genetic markers than found on the total immunoglobulin of the same individual (Allen et al., 1964 Yount et al., 1968). 

Electrophoretic separations of solutes are determined primarily by the mass-to-charge ratio of the solute. However, certain electrophoretic separations use gel or paper supports for the electrolyte, and adsorptive and molecular-size fractionations may be as significant as electrophoretic fractionation in these systems. Hall (1970) found that polyacrylamide gel electrophoresis of aquatic humic substances gave size fractionations corresponding to Sepha-dex gel fractionation however, individual fractions were poorly resolved in polyacrylamide gel electrophoresis. Clapp (1957) and Mortensen and Schwendinger (1963) used electrophoresis with electrolyte supporting curtains to separate colorless polysaccharides from humic substances in soil-water extracts. 

The third component in the Wzy/Wzx-dependent pathway is the Wzz protein. Wzz generates the strain-specific chain length distribution of 0 polysaccharide chain lengths as reflected in characteristic clusters of bands following gel electrophoresis of LPS samples.The function of the Wzz protein is not required for bacterial growth in the laboratory setting, as polymerization can proceed in the absence of this protein. However, the distribution of the 0 polysaccharide chain length is critical for virulence. 

Trichloropyrimidinyl and chlorotriazinyl dye derivatives of polysaccharides are considered to be more satisfactory than Ramazol dye derivatives, the ether linkage being considered the weakest of those present. The dyeing of polysaccharides has been advocated as an aid to their identification in gel electrophoresis, gel filtration, and cellulose acetate electrophoresis, although it must be borne in mind that such deriva-tizations alter the characteristics of the polysaccharide under examination. 

Li, J., Wang, Z. and Altman, E. In-source fragmentation and analysis of polysaccharides by capillary electrophoresis/mass spectrometry. Rapid Commun Mass Spectrom, 19,1305, 2005. 

Wang, Q., Ding, F., Zhu, N., He, R. and Fang, Y. Determination of the compositions of polysaccharides from Chinese herbs by capillary zone electrophoresis with amperometric detection. Biomed Chromatogr, 17, 483, 2003. 

Mistry, K., Krulll, L, and Grinberg, N., Size-exclusion capillary electrochromatographic separation of polysaccharides using polymeric stationary phases. Electrophoresis, 24, 1753, 2003. 

Alginate lyase from Turbo cornutus acted on sodium alginate and resulted in a rapid decrease in viscosity and a gradual increase in reducing power of the substrate solution. The result of gel filtration of the reaction products indicated that uronic acid oligosaccharides were liberated as the final product. The fact that the enzyme attacked preferentially the D-mannuronate-rich moieties of the alginate molecule was demonstrated by an analysis with paper electrophoresis of the reaction products after acid treatment, which separated the uronic acid oligosaccharides from the uronic acid polysaccharides. 

FIGURE 1. Affinity electrophoresis of a pea leaflet extract. Separation gels contained varying concentrations of a polysaccharide fraction isolated from cotyledons. For quantification, an aliquot of the polysaccharide fraction was subjected to acid hydrolysis and the monosaccharide content was monitored. Cytosolic isozyme I Plastidic isozymes II and III... 

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