Glycosylation separation

Human interleukin 2, a 133-residue protein, has been separated into multiple molecular forms by selective immunoaffinity chromatography and chromatofocusing. Most of the heterogeneity has been attributed to variations in glycosylation of the threonine residue in position 3 of the polypep-... 

Supercritical fluid extraction (SEE) is another modern separation technology usually employed to extract lipophilic compounds such as cranberry seed oil, lycopene, coumarins, and other seed oils. Anthocyanins generally and glycosylated anthocyanins in particular were considered unsuitable for SEE due to their hydrophilic properties, since SEE is applicable for non-polar analytes. However, a small amount of methanol was applied as co-solvent to increase CO2 polarity in anthocyanin extraction from grape pomace. New applications of SEE for anthocyanin purification have been reported for cosmetic applications from red fruits. ... 

Glycosyl-residue compositions were determined by GC-CIMS analysis of the per-O-trimethylsilylated methyl glycosides [23] separated on a fused-silicaDB-1 capillary column. The identity of each peak was confirmed by CIMS on a HP-5989 MS Engine (Hewlett-Packard). 

The affinity chromatography on ConA - cellulose indicated the presence of small N-glycosylation of all forms of exopolygalacturonases present in carrot roots (unpublished results). This method was usefull for purification of these enzymes from other protein inpurities but was completely uneffective by separation of individual forms (Fig. 4). 

Carbohydrates play a major role in protein bioactivity, bioavailability, and antigenicity therefore, the understanding of the glycosylation of protein molecules is very important in the development of effective glycoprotein therapeutics.172 In recent years, there has been considerable activity in the development of simple, rapid, and reliable separation methods for the analysis of... 

C13HuC1N307 2,2 -Anhydro-[5-chloro-l-(3,5-di-0-acetyl-/ -D-arabino-furanosyl)-6-oxocytosine] (ACAFCC)166 I4t Z — 8 D = 1.56 R = 0.066 for 818 intensities. The glycosyl disposition of the anhydronucleoside is constrained to the syn (— 68.8°) orientation. The conformation of the D-arabinofuranosyl group is a flattened 4E (232.6°, 18.0°), and the exocyclic, C-4 -C-5 bond torsion-angle is g auche+ (50.2°). The adjacent bases are connected by N-H O hydrogen-bonds between the N-4 atom of one molecule and the carbonyl oxygen atom (0-4) of another. The twofold-symmetry-related bases are stacked, with an interbase separation of 329 pm. 

These model reactions were of great value in the extension of the C-glycosylation reaction with malonic esters to five-membered ring-systems. Treatment of 2,3 5,6-di-O-isopropylidene-a-D-manno-furanosyl bromide122,123 (150) with diethyl sodiomalonate led to an anomeric mixture of C-glycosyl compounds that could be separated by column chromatography, with the a (151) and /B (152) anomers in... 

Fig. 1. General microreactor for organic transformations as the round-bottom flask equivalent. The microreactor incorporates (i) a laminar flow mixing section of separate streams, exemplified for glycosylation, (ii) reaction channel, inlet for quench, and an outlet (Ratner et al. 2005 Murphy 2006)...

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