Citric Acid → HCID 131724567, compound

Participants

Reaction Steps

1-thio-b-galactopyranose (1.5 mg) is dissolved in 150 μl of buffer A (20 mM citric acid, 35 mM Na2HPO4, 108 mM NaCl, 1 mM ethylenediamine-tetraacetic acid (EDTA)) and 150 μl of ethanol, N-(m-(3-trifluoromethyl)diazirine-3-yl)phenyl)-4-maleimido-butyramide (MAD, 3.3 mg, preparation according to A. Collioud et al., Bioconjugate Chemistry 4, 528-536 (1993)) is dissolved in 150 μl of ethanol and an additional 150 μl of buffer A. Both solutions are mixed. The mixture is stirred and allowed to react at room temperature for 90 min, to give the compound MAD-Gal. The product is purified by HPLC. The HPLC flow rate is 2 ml/min. For gradient elution (0 min.-100% A; 5 min.-100% A; 20 min.-70% A; 30 min.-0% A; 40 min. 0%A), solvent B is added to solvent A (solvent A, 0.1% TFA in water; solvent B, 0.1% TFA in acetonitrile/water (4:1 by volume)). Detection is performed at 254 nm and fractions of the eluant are collected. 13C.NMR (D2O chemical shift (δ) in parts per million): 179.14, 178.40, 177.99, 177.86, 173.30, 137.79, 129.53, 129.38, 122.46, 121.84, 118.32, 84.42, 84.27, 78.99, 78.91, 73.84, 73.66, 69.73, 69.68, 68.58, 68.49, 60.80, 60.69, 39.61, 38.66, 38.46, 37.71, 35.83, 33.57, 22.68, 22.57 ppm. Mass Calc (M−H): 561.1 Da/e, found: 561.1 Da/e.

Provenance & Evidence