Carbohydrate arrays

In a related study, Turnbull and coworkers described the attachment of carbohydrates to amino-derivatized glass slides. They found a significant rate enhancement when performing this step under microwave irradiation as compared to classical heating [47]. This method should be an efficient aid for the construction of functional carbohydrate array systems. 

Frequently, integral membrane proteins are glycosylated with complex carbohydrate arrays. Explain how glycosylation further enhances the asymmetrical orientation of integral proteins. 

The staining procedure for carbohydrate microarrays is basically identical to the routine procedure for immunohistology. Immunostaining steps of carbohydrate arrays are listed below (see Notes 4 and 6 and Fig. 3). 

Houseman, B. T. and Mrksich, M. (2002) Carbohydrate arrays for the evaluation of protein binding and enzymatic modification. Chem. Biol. 9(4), 443-454. 

In recent years, different formats of biochips such as protein arrays, peptide arrays, antibody arrays, living cell arrays, tissue arrays, and carbohydrates arrays have been developed (Figure 7.1). They are associated with different applications and show specific advantages as well as definite bottlenecks. 

Both Siegel et al. (122) and Lawrence et al. (123) have described automated systems for the purification of small arrays of amines and amides. A 48-member array of P-amino alcohols prepared from epoxides and amines was purified using SPE by Shuker et al. (124). Blackburn et al. (125) have described the purification of a 60-member 3-aminoimidazo[l,2-fl]pyridine array obtained from a multiple-component condensation, and Bussolari et al. (126) purified a small array of phenylpropyl amines obtained from dihydrocoumarins and amines. A few applications where ion-exchange resins have been substituted with other solid phases have also recently appeared. For example, the purification of several carbohydrate arrays tagged as hydrophobic O-laurates using Ci8 silica producing up to 10-30 mg of >90% pure individuals was described by Nilsson et al. (127), and Curran et al. purified fluorous-... 

Schwarz M, Spector L, Gargir A, Shtevi A, Gortler M, Alt-stock RT, Dukler AA, Dotan N. A new kind of carbohydrate array, its use for profihng antiglycan antibodies, and the discovery of a novel human cellulose-binding antibody. Glycobiology 2003 13 749-754. 

Biskup MB, Muller JU, Weingart R, Schmidt RR. New methods for the generation of carbohydrate arrays on glass slides and their evaluation. Chembiochem 2005 6 1007-1015. 

Manimala J, Li Z, Jain A, VedBrat S, Gildersleeve JC. Carbohydrate array analysis of anti-Tn antibodies and lectins reveals nnexpected specificities implications for diagnostic and vaccine development. ChemBioChem 2005 6 2229-2241. 

Shirahata N, Hozumi A, Miura Y, Kobayashi K, Sakka Y, Yonezawa T. An efficient matrix that resists the nonspecific adsorption of protein to fabricate carbohydrate arrays on silicon. 

Lawrie CH, Marafioti T, Hatton CSR, Dirnhofer S, Roncador G, Went P, Tzankov A, Pileri SA, Pulford K, Banham AH. Cancer-associated carbohydrate identification in Hodgkin s lymphoma by carbohydrate array profiling. Int. J. Cancer 2006 118 3161-3166. 

Finally, hydroquinone was connected with its C2 via a pentaethylene glycol Hnker to long alkyl chains with terminal thiol functions. The latter self-assembled on a gold surface to give monolayers. Upon oxidation the hydroquinone was oxidized to quinone and could perform a Diels-Alder reaction with cyclopentadienyle-thanol-conjugated carbohydrate analogs. This set-up is used in chip-based carbohydrate arrays for the evaluation of protein binding and modification [268]. 

Fig. 4-3b). Reaction of azides and alkynes in the presence of catalytic Cul proceeded regioselectively in 8 hours to give a 1,3-dipolar cycloaddition product (i.e., 1,4-disubstituted 1,2,3-triazole) in good yield, whereas thermal cycloaddition gave a mixture of 1,4- and 1,5-disubstituted 1,2,3-triazole. The Cu(I)-catalyzed reaction was applied successfully to fabricate carbohydrate arrays in microtiter plates [45-47]. 

Shin I, Cho J W, Boo D W (2004). Carbohydrate arrays for functional studies of carbohydrates. Combinat. Chem. High Throughput Screen. 7 565-574. 

Shin, L, Cho, J. W., and Boo, D. W. (2004) Carbohydrate arrays for functional studies of carbohydrates Comb Chem High Throughput Screening 7, 565-74. 

In order to make the best use of available technologies of carbohydrate microarrays, it is important to conceptually understand the design and chemical principles of different carbohydrate microarray platforms. Different platforms may be technically complementary and can be applied in combination in addressing biomedical questions. For this purpose, we outhne below a number of carbohydrate array platforms based on the chemical principle of array construction. These include technologies that directly utilize underivatized carbohydrates in microarray construction, technologies that require chemical modification of carbohydrates... 

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