Oligosaccharides method

Cummings, R. D., Merkle, R. A., and Stults, N. L. (1989). Separation and Analysis of Glycoprotein Oligosaccharides. Methods Cell Biol 32 141. 

R406 M. Kitagawa, K. Yamashina, S. Tojyo and D. Ulam-Orgikh, In Search for Molecules which Calculate Quantum Mechanically , Mem. Inst. Sci. Ind. Res., Osaka Univ., 2000, 57, 96 R407 A. Klein, G. Strecker, G. Lamblin and P. Roussel, Structural Analysis of Mucin - Type O - Linked Oligosaccharides , Methods Mol. Biol. (Totowa, N.J.), 2000,125,191... 

H. M. Flowers. Chemical synthesis of oligosaccharides, Methods Enzymol., 50 (Complex Carbohydrates, Part C) (1978) 93-121. 

Dell A. Preparation and desorption mass spectrometry of permethyl and per-acetyl derivatives of oligosaccharides. Methods Enzymol 1990 193 647-660. 

Takasaki, et al. (1982). Hydrazinolysis of Asparagine-linked Sugar Chains to Produce Free Oligosaccharides, Methods Enzymol. 83 263 268. 

Takasaki, S. Misuochi, T. Kobaia, A. Hydrazinolysis of asparagine-linked sugar chains to produce free oligosaccharides. Methods Enzymol. 1982, 83, 263-268. 

A central theme of carbohydrate chemistry today is the construction of oligosaccharides as they are found in glycoproteins (H. Paulsen, 1990). Three closely related methods have proven to be reliable in complex syntheses, namely the halide, imidate, and thioglycoside methods. 

One widely used method involving protected compounds is solid-phase synthesis (polymer-supponed reagents). This method has the advantage of requiring only a simple workup by filtration such as in automated syntheses, especially of polypeptides, oligonucleotides, and oligosaccharides. 

Note. This prefix does not include the oxygen of the glycosidic group. This is the appropriate method for naming natural products if the trivial name includes the OH group. The system is also used to name oligosaccharides (see 2-Carb-37). 

There are two established methods in use for citing locants either in parentheses between the glycosyl and glycose terms, or in front of the glycosyl prefix, as in the names of glycosides. The former (preferred) method is derived from that used to designate residues in oligosaccharides (see 2-Carb-37 and -38). 

I. General Introduction Glycosidation Methods for Oligosaccharide Synthesis. .. 178... 

Of the many methods described for glycoside synthesis, those predominantly used in current oligosaccharide synthesis, in which the glycosyl donor has O- or N- functionality at C-2 (that is, excluding 2-deoxyglycosides), are the following ... 

To allow all culture productiou to be coutrolled, a method for rapid analysis is required. Prior to development of an LC-MS method, the analysis was both complex and time-consuming, involving the purification of a relatively large amount of the antibody using affinity chromatography, enzymatic release, and subsequent derivatizafion of the oligosaccharides and their analysis by using capillary electrophoresis. 

Figure 5.9 Principal oligosaccharide structures found on recombinant rituximab. Reprinted from J. Chromatogr., A, 913, Wan, H. Z., Kaneshiro, S., Frenz, J. and Cacia, J., Rapid method for monitoring galactosylation levels dnring recombinant antibody production by electrospray mass spectrometry with selective-ion monitoring , 437-446, Copyright (2001), with permission from Elsevier Science.

In a study completed during the early development of f.a.b.-m.s., both f.d. and f.a.b. were used to characterize 101 fractions containing neutral oligosaccharides isolated from human milk. Samples were examined as their peracetylated alditols. In subsequent work, the structures of two minor acidic oligosaccharides from human milk were investigated. The per-methylated derivatives were analyzed by f.a.b.-m.s., and their compositions and sequences were defined by the f.a.b. data. Methylation analysis and partial formolysis were the other principal methods used. 

Plant material fractionation. The detailed steps of isolation and separation of oligosaccharide fractions were described earlier [10]. Pectin was separated by boiling the cell walls in 0.5 M ammonium-oxalate buffer, pH 5.2 at 100 C for Ih. The dialyzed solution of pectin was hydrolyzed with 0.15M HCl for 3h at 100 C. Neutralized and desalted hydrolysate was loaded to the column (lx90cm) filled with biogel TSK HW-40 (Toyo Soda, Japan) equilibrated and eluted with 50mM sodium acetate (pH 5.2) at a rate of 0.3ml/min. In all fractions (1ml) the sugars were determined by o-toluidine method (Resnikov et al., 1982) and fraction IP was collected as shown on Figure 1. 

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