环氧氯丙烷活化琼脂糖凝胶过程强化及性能评价

Process Intensification and Characterization of Agarose Gel Activation with Epichlorohydrin

环氧氯丙烷的低水溶性和活化过程中环氧基水解制约了高活化效率的获得.通过引入亲水性有机试剂二甲基亚砜,研究了提高Sepharose CL-6B介质环氧基活化密度的方法.研究结果显示,二甲基亚砜溶剂体系消除了琼脂糖凝胶与环氧氯丙烷之间的相界面,形成了均相反应体系,强化了活化反应.在10%(φ)环氧氯丙烷、0.8mol/L氢氧化钠及反应时间2.5h的优化条件下,环氧基活化密度可达100μmol/mL以上.上述活化过程对介质的性能没有明显影响.活化介质对5-氨基吲哚的偶联密度达到95.3μmol/mL,较常规活化方法提高75%以上.

The epoxy density of agarose gels is generally limited by low water-solubility of epichlorohydrin （ECH） and its hydrolysis during the activation process. In this work, a hydrophilic organic agent, dimethyl sulfoxide （DMSO）, was introduced to enhance the epoxy activation of agarose gel. The results showed that the phase interface of epichlorohydrin and agarose gel suspension was diminished in the DMSO solution. Thus the epoxy activation was enhanced by a high ECH solubility in DMSO. An optimal epoxy density over 100 μmol/mL was achieved at 10%（φ） epichlorohydrin and 0.8 mol/L sodium hydroxide. The activated gel did not show any loss in performance. The coupling density of a ligand, 5-aminoindole, on the gel reached 95.3 μmol/mL, being 75% higher than that obtained with common epoxy-activated agarose gel. It exhibited intriguing potential in the preparation of functional agarose gel with a high density of ligand.