亲和层析固定相的制备及应用研究  

Study on preparation and application of the stationary phase for affinity chromatography

作　　者：莫旖[1] 柳畅先[1] 

机构地区：[1]中南民族大学化学与材料科学学院,分析化学国家民委重点实验室,湖北武汉430074

出　　处：《化学研究与应用》2013年第4期457-461,共5页Chemical Research and Application

基　　金：中南民族大学自然科学基金项目(XTZ09005)资助

摘　　要：以壳聚糖为载体,戊二醛为交联剂,氨基三乙酸(NTA)为配基,制备固载金属离子的亲和吸附剂,试验了交联剂浓度、配基浓度、偶联温度及时间对配体固载量的影响,最佳条件为戊二醛浓度0.55%、NTA浓度0.4mol·L-1、偶联温度65℃、偶联时间10h。层析分离纯化豆类脲酶,锌柱和镍柱的酶回收率分别为88.5%和80.4%,纯化倍数分别为3.7和6.1,实验结果表明镍柱对脲酶有较高的分离纯化选择性。Chitosan was used as support, glutaraldehyde as cresslinking agent, nitrilotriacetic acid（NTA） as the chelating ligand. The affinity adsorbent was prepared by immobilized metal ions. The effects on binding amount of ligand of cress-linking agent concentration,genin concentration, binding temperature and time were tested. The optimum condition of the concentration of glutaraldehyde and NTA were 0. 55% and 0. 4mol ·L^-1 respectively, the binding temperature and time were 65℃ and 10h respectively. Urease from beans was purified by the affinity chromatography. The recovery rate of urease were 88. 5% and 80. 4%, the purification fold were 3.7 and 6. 1 with column chelating Zn（ Ⅱ ）and Ni（ Ⅱ ）respectively. The results showed that affinity chromatography with column chelating Ni （Ⅱ） had a higher selectivity for separation and purification of urease.

关 键 词：亲和层析 脲酶 壳聚糖 

分 类 号：O658[理学—分析化学]