Method for preparing water-soluble affinity ultrafiltration carrier
A water-soluble and affinity technology, applied in ultrafiltration and other directions, can solve the problem of difficult separation of components with similar molecular weights, and achieve the effect of fast reaction speed, short time and good purification effect.
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Embodiment 1
[0015] Put 1 gram of dextran with a molecular weight of 2 million purchased from Sigma into a beaker, add 15 mL of water, and disperse evenly with a magnetic stirrer to completely dissolve the dextran, then add 0.4 gram of sodium hydroxide, add 10 mL of water, and stir with a magnetic force. Stir for 35 minutes; add 1 mL of epichlorohydrin, and quickly magnetically stir to disperse evenly; quickly move the above-mentioned stirring solution to a constant temperature shaker at 57±2°C, the shaker rotates at 100~110r / min, and react in the dark for 1.8 hours; The above reaction liquid is transferred to an ultrafiltration cup, the membrane molecular weight cut-off is 100,000, and the remaining sodium hydroxide and epichlorohydrin small molecules are removed. First wash with distilled water to pH 8.0 ± 0.2, and the retentate is 40 ± 5 mL, and then wash with 0.1 M, pH 9.5 sodium carbonate buffer 100 ± 5 mL to the retentate to 30 ± 2.5 mL, the entire process requires Protect from light; di...
Embodiment 2
[0018] Weigh 3 grams of dextran with a molecular weight of 2 million purchased from Pharmacia Company into a beaker, add 50 mL of water, disperse evenly with a magnetic stirrer, and completely dissolve the dextran, then add 1.3 grams of sodium hydroxide and 15 mL of water. Stir with a magnetic stirrer for 30 minutes; then add 3±0.2 mL of epichlorohydrin, and quickly stir and disperse evenly by magnetic stirring; quickly move the above stirring liquid to a constant temperature shaker at a temperature of 58±2°C, and a shaker speed of 100~110r / min , Dark reaction for 2 hours; transfer the above reaction solution to an ultrafiltration cup, the membrane molecular weight cut-off is 100,000, remove the remaining sodium hydroxide, epichlorohydrin small molecules, first wash with distilled water to pH 8.0 ± 0.2, And until the retentate is 80±5mL, then use 0.1M, pH9.5 sodium carbonate buffer 300±5mL to wash to the retentate volume of 80±5mL, the whole process needs to be protected from ligh...
Embodiment 3
[0021] Put 1.5 grams of dextran with a molecular weight of 2 million purchased from Pharmacia into a beaker, add 25 mL of water, disperse evenly with a magnetic stirrer, and completely dissolve the dextran, then add 0.6 grams of sodium hydroxide, and then add 8 mL of water. Stir with a magnetic stirrer for 40 minutes; then add 1.5 mL of epichlorohydrin, and magnetically stir to quickly disperse it evenly; quickly move the above stirring liquid to a constant temperature shaker at a temperature of 58±2°C, and a shaker speed of 100~110r / min. Keep the reaction in the dark for 1.9 hours; transfer the above reaction solution to an ultrafiltration cup, the membrane has a molecular weight cut-off of 100,000, remove the remaining sodium hydroxide and epichlorohydrin small molecules, first wash with distilled water to pH 7.8 ± 0.2, and Until the retentate is 40±5mL, then wash with 150±5mL of 0.1M, pH9.5 sodium carbonate buffer solution to a volume of 40±2mL. The whole process needs to be pr...
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