壳聚糖金属螯合亲和吸附剂的制备

Preparation of immobilized metal ion affinity adsorbent for recombinant human-like collagen adsorption

目的研究利用壳聚糖为载体、戊二醛为交联剂、亚氨基二乙酸(IDA)为螯合配基制备金属螯合亲和吸附剂的条件。方法戊二醛对壳聚糖进行交联,环氧氯丙烷法进行载体活化和螯合配基的偶联。键合IDA后与Cu2+螯合。结果采用0.22 mL/g(壳聚糖)戊二醛对壳聚糖进行交联。交联壳聚糖加入0.4 mol/L NaOH与体积分数为0.5的二甲亚砜的混合溶液中,充分搅匀,25℃下缓慢滴加1.2 mL(体积分数为0.3)的环氧氯丙烷,升温至40℃,170 r/min下振荡反应3h进行活化。之后,以4.26 g/g(壳聚糖)浓度的IDA进行连接反应,65℃下反应18 h。结论采用优化条件制备的金属螯合亲和吸附剂对Cu2+有较大螯合量,可用作固定化金属亲和层析的固定相。

Aim To study the preparation conditions of the metal chelate gel with chitosan as support. Methods Chitosan was crosslinked with glutaraldehyde. Crosslinking support was activated with epiehlorohydrin. Then the gel was prepared with iminodiacetic acid （IDA） as chelating ligand and Cu^2＋ as chelating metal ion. Results The adsorbents （0. 22 mL/g（chitosan） mass fraction of glutaraldehyde） had good stability and mechanical strength. The epoxy density was the most with 1：1 NaOH（0. 4 tool/L） and 50% DMSO as activating accelerator for 3 h. The study on the linkaging of IDA demonstrates that the adsorbents synthesized with 4.26 g/g （chitosan） IDA, at 65℃ for 18 h could chelate copper ion the most. Conclusion The metal chelate affinity adsorbent prepared with optimized conditions could well chelate Cu^2＋ and could be used in immobilized metal ion affinity chromatography as stationary phase.