Expanded bed adsorption medium with sulfhydryl ethylpyridine and sulfone group as ligand separation antibody and preparation method
A technology of mercaptoethyl pyridine and adsorption medium, which is applied in the field of expanded bed adsorption and separation of proteins in chromatographic separation, can solve the problems of low antibody selectivity and no emphasis on thiophilic effect.
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Embodiment 1
[0015] Stir 50g of cellulose viscose and 40g of tungsten carbide powder in a 500ml round-bottomed flask at 300rpm, add 300g of pump oil, rotate at 700rpm, inversely suspend and regenerate to make balls, wash with boiling water and deionized water for 3 to 5 times, Screen cellulose / inorganic weighting agent composite microspheres with a particle size of 50-250 μm as the matrix; 10ml of drained composite microspheres, 3.6ml of divinyl sulfone, 9ml of 0.25M pH12 sodium carbonate buffer and 1.5ml of bismuth Methyl sulfoxide was added into the Erlenmeyer flask, activated in a shaker at 180 rpm at 25°C for 4 hours, the mixed solution was removed by suction filtration, washed with deionized water, and sucked dry. Add 4-mercaptoethylpyridine with 3 times the molar amount of double bonds and 10ml of 0.5M sodium hydroxide solution containing 25mg / ml ammonium persulfate, perform coupling reaction in a shaker at 180rpm at 60°C for 8 hours, and remove the mixed solution by suction filtratio...
Embodiment 2
[0017] Stir 50g of cellulose viscose and 40g of tungsten carbide powder in a 500ml round-bottomed flask at 300rpm, add 300g of pump oil, rotate at 700rpm, inversely suspend and regenerate to make balls, wash with boiling water and deionized water for 3 to 5 times, Screen cellulose / inorganic weighting agent composite microspheres with a particle size of 50-250 μm as the matrix; 10ml of drained composite microspheres, 0.9ml of divinyl sulfone, 9ml of 0.2M pH12 sodium carbonate buffer and 1.5ml of bismuth Methyl sulfoxide was added into the Erlenmeyer flask, activated in a shaker at 180 rpm at 25°C for 4 hours, the mixed solution was removed by suction filtration, washed with deionized water, and sucked dry. Add 4-mercaptoethylpyridine with 3 times the molar amount of double bonds and 10ml of 0.8M sodium hydroxide solution containing 25mg / ml ammonium persulfate, perform coupling reaction in a shaker at 180rpm at 60°C for 8 hours, and remove the mixed solution by suction filtration...
Embodiment 3
[0019] Stir 50g of cellulose viscose and 80g of tungsten carbide powder in a 500ml round-bottomed flask at 300rpm, add 310g of pump oil, rotate at 800rpm, inversely suspend and thermally regenerate to make balls, wash with boiling water and deionized water for 3 to 5 times, Screen cellulose / inorganic weighting agent composite microspheres with a particle size of 50-250 μm as the matrix; 10ml of drained composite microspheres, 1.8ml of divinyl sulfone, 9ml of 0.5M pH12 sodium carbonate buffer and 1.5ml of di Methyl sulfoxide was added into the Erlenmeyer flask, activated in a shaker at 180 rpm at 25°C for 4 hours, the mixed solution was removed by suction filtration, washed with deionized water, and sucked dry. Add 4-mercaptoethylpyridine with 3 times the molar amount of double bonds and 10ml of 0.3M sodium hydroxide solution containing 25mg / ml ammonium persulfate, perform coupling reaction in a shaker at 180rpm at 60°C for 8 hours, and remove the mixed solution by suction filtr...
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