无水体系中环氧氯丙烷活化琼脂糖凝胶制备高载量固定化金属亲和层析介质

在以二甲基亚砜为溶剂的无水体系中,利用环氧氯丙烷对琼脂糖凝胶Sepharose 6 Fast Flow进行活化,并偶联亚氨基二乙酸和Cu2+制备了固定化金属亲和层析介质. 结果表明,该体系中环氧氯丙烷对琼脂糖凝胶的活化效率大幅度提高,在40%(j)环氧氯丙烷、0.02 g/mL NaOH及50℃、反应时间4 h的优化条件下,环氧基活化密度最高达165 mmol/mL,较目前报道的最高值提高50%以上. 最终所制介质的Cu2+螯合密度为128.3 mmol/mL,对BSA的平衡吸附容量达2.05 mmol/L. 以0.5 mol/L咪唑为洗脱剂,被吸附的BSA洗脱率可达90%以上.

Activation of Sepharose with Epichlorohydrin in Water-free System for Preparation of High Capacity Immobilized Metal Ion Affinity Adsorbent

Epoxy activation of agarose gel Sepharose 6 FF by epichlorohydrin (ECH) was carried out in a water-free medium with dimethyl sulfoxide as solvent. The activated agarose gel was subsequently coupled with iminodiacetic acid and Cu2+ ions for preparation of immobilized metal ions affinity chromatographic (IMAC) adsorbent. The results indicated that the epoxy activation efficiency was significantly enhanced in the water-free system, such that a epoxy density of agarose gels as high as 165 mmol/mL was achieved at the optimized conditions of 40%(j) ECH, 0.02 g/mL NaOH, 50℃ and 4 h, which is about 50% higher than the highest value reported so far. The prepared IMAC adsorbent showed a high chelating capacity for Cu2+ ion up to 128.3 mol/mL, thus it ensured a high equilibrium adsorption capacity of 2.05 mmol/L for bovine serum albumin, and 90% of the adsorbed protein could be efficiently eluted when 0.5 mol/L imidazole was used as elute.