摘要
固定化金属离子亲和层析(IMAC)在生物分离中得到广泛的应用,但介质存在着较强的非特异性吸附,导致亲和介质分离效率下降。为减少IMAC的非特异性吸附,选用聚乙二醇单甲醚5000(m PEG 5000)为原料,经醛基化后修饰磁性壳聚糖微球固定化Cu2+亲和介质(MCS@Cu2+),得到m PEG修饰固定化Cu2+亲和介质(m PEG@MCS@Cu2+)。以牛血清白蛋白(BSA)作为大分子模型,以修饰的亲和介质对BSA吸附阻拒率为指标,优化了m PEG修饰MCS@Cu2+的工艺条件。结果表明:在反应温度为40℃、反应时间为20 h、n-CHO:n-NH2为4:1时,在m PEG@MCS@Cu2+上修饰的m PEG含量最大为12.5%,与MCS@Cu2+相比,修饰介质对BSA吸附阻拒率可达到37.4%。说明经m PEG修饰的MCS@Cu2+亲和介质在应用于富集活性分子时,可有效减少大分子非特异性吸附,提高IMAC分离效率。
Immobilized metal ion affinity chromatography (IMAC) was widely used in biological separation. The phenomenon about strong non-specific absorption of large molecule could not be ignored. In order to improve it, affinity media-MCS@Cu2+ was modified by mPEG 5000 to produce modified affinity media- mPEG@MCS@Cu2+. The BSA was used as the model large molecule and the resistance rejection rate of BSA as standard, the optimal procession conditions about the modified affinity media were as follows: reaction temperature was 40 ℃ ; reaction time was 20 h, molar ratio of -CHO:-NH2 was 4:1. The content of mPEG was 12.5%. Compared with MCS@Cu2+, resistance rejection rate about BSA was 37.4% finally for mPEG@MCS@Cu2+. It revealed that mPEG@MCS@Cu2+ can effectively reduce the non-specific absorption and improve the efficiency of IMAC, when it was used to separate small molecules.
出处
《食品科技》
CAS
北大核心
2015年第10期221-224,共4页
Food Science and Technology
基金
国家自然科学基金项目(51372043)
广西大学区级大学生创新项目(201410593100)