* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.
methyl (2-amino-2-deoxy-3,4-di-O-methyl-6-O-sulfonato-α-D-glucopyranosyl)-(1>4)-(2,3-di-O-methyl-β-D-glucopyranosyluronic acid)-(1>4)-(2,3,6-tri-O-sulfonato-α-D-glucopyranosyl)-(1>4)-(2,3-di-O-methyl-α-L-idopyranosyluronic acid)-(1>4)-2,3,6-tri-O-sulfonato-α-D-glucopyranoside sodium salt[ No CAS ]
[ 756526-03-1 ]
C55H81N2O58S7(9-)*9Na(1+)[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
45%
With sodium hydrogencarbonate In water at 20℃; for 16h;
To a dry, stirred solution of a pentasaccharide (20 mg, 0.013 mmol), shown above, in 0.5% sodium hydrogen carbonate (2 mL) is added Bis(NHS)PEO5 (Pierce Chemical Company) (7 mg, 1.1 equiv.). After stirring for 16 hours at room temperature, the reaction mixture was deposited on a column Sephadex G-25F, which was eluted with sodium chloride. The fractions comprising the product were concentrated and desalting was carried out on the same column, which was eluted with water. After lyophilizing, 11 mg of NHS-PEGylated pentasaccharide 9 (45%) was obtained. Mass: ES1 method, negative mod: chemical mass = 1922; experimental mass = 1923.
Procedure for conjugation of saccharide 3 with HSA
General procedure: N-hydroxydisuccinimidyl (BS) PEG5 was purchased from Pierce. The sugar ( 10 μmol ) dissolved in DMSO (250 μL) containing triethylamine (25 eq), was added drop-wise to a mixture of BS(PEG)5 ( 10 eq) in DMSO (250 μL). After 3 hours under vigorous stirring, the activated oligosaccharide was precipitated by addition of nine volumes (9 mL) of ethyl acetate. The pellet obtained by subsequent centrifugation was washed with ethyl acetate (10 times× 3 mL), and freeze dried. After spectrophotometric determination of active ester groups, the pellet was incubated overnight with the protein in 100 mM NaPi pH 7.2 at an activated glycan:protein ratio of50:1 for compounds 18 and 19, and 75:1 for compound 20.
bis-sodium 3-aminopropyl 1-O-[2-acetamido-2-deoxy-α-D-glucopyranosyl phosphate 4-(2-acetamido-2-deoxy-α-D-glucopyranosyl phosphate)][ No CAS ]
C37H62N4O27P2(2-)*2Na(1+)[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
With triethylamine In dimethyl sulfoxide for 3h;
Procedure for conjugation of saccharide 3 with HSA
General procedure: N-hydroxydisuccinimidyl (BS) PEG5 was purchased from Pierce. The sugar ( 10 μmol ) dissolved in DMSO (250 μL) containing triethylamine (25 eq), was added drop-wise to a mixture of BS(PEG)5 ( 10 eq) in DMSO (250 μL). After 3 hours under vigorous stirring, the activated oligosaccharide was precipitated by addition of nine volumes (9 mL) of ethyl acetate. The pellet obtained by subsequent centrifugation was washed with ethyl acetate (10 times× 3 mL), and freeze dried. After spectrophotometric determination of active ester groups, the pellet was incubated overnight with the protein in 100 mM NaPi pH 7.2 at an activated glycan:protein ratio of50:1 for compounds 18 and 19, and 75:1 for compound 20.
Procedure for conjugation of saccharide 3 with HSA
General procedure: N-hydroxydisuccinimidyl (BS) PEG5 was purchased from Pierce. The sugar ( 10 μmol ) dissolved in DMSO (250 μL) containing triethylamine (25 eq), was added drop-wise to a mixture of BS(PEG)5 ( 10 eq) in DMSO (250 μL). After 3 hours under vigorous stirring, the activated oligosaccharide was precipitated by addition of nine volumes (9 mL) of ethyl acetate. The pellet obtained by subsequent centrifugation was washed with ethyl acetate (10 times× 3 mL), and freeze dried. After spectrophotometric determination of active ester groups, the pellet was incubated overnight with the protein in 100 mM NaPi pH 7.2 at an activated glycan:protein ratio of50:1 for compounds 18 and 19, and 75:1 for compound 20.