一种新型金属螯合亲和色谱固定相的制备及评价  

Preparation and evaluation of a novel solid phase in immobilized metal affinity chromatography(IMAC)

作　　者：骆红琴[1] 李乃瑄[1] 孙文晓[2] 

机构地区：[1]天津理工大学化学化工学院,天津300191 [2]华东理工大学材料科学与工程学院,上海200237

出　　处：《天津理工大学学报》2007年第2期59-62,共4页Journal of Tianjin University of Technology

摘　　要：采用溶胶-凝胶法,在正庚烷乳液中生成1.5μm的无孔脲醛树脂-二氧化锆复合微球基体,其与丙烯酸甲酯进行迈克尔加成反应后,通过亲核取代反应与螯合剂亚氨基二乙酸(IDA)键合,制成IDA-UF-ZrO2固定相,此固定相与铜离子(Ⅱ)螯合后,形成金属螯合亲和色谱固定相(Cu-IDA-UF-ZrO2).以牛血清白蛋白(BSA)为样品,考察了两种新合成的固定相的色谱性能,结果表明BSA在Cu-IDA-UF-ZrO2上的保留时间比在IDA-UF-ZrO2上的保留时间有所增加,同时BSA在Cu-IDA-UF-ZrO2上的保留主要受固定金属离子与蛋白质侧链上氨基酸间配位作用的影响.1.5 μm nonporous Urea-Formaldehyde resin-Zirconia composite microspheres（ UF-ZrO2 ） as the matrix were prepared in normal heptane latex by sol-gel method, firstly activated by Michael Addition Reaction with methacrylate, and subsequent bonded with chelating ligand iminodiacetic acid（IDA） by nucleophilic substitution, yielded a intermediate product IDA-UF-ZrO2. When IDA-UF-ZrO2 was chelated with copper ion （ Ⅱ ）, a novel solid phase in IMAC was composed. Using bovine serum albumin （BSA） as testing sample, the chromatography perfor-mance of the two newly synthesized solid phase was tested , the result indicated that the retension time of BSA on Cu-IDA-UF-ZrO2 was longer than that on IDA-UF-ZrO2, also the retension of BSA on Cu-IDA-UF-ZrO2 was mainly dominated by the chelation of the immobilized metal ion and amino acid on the side chain of BSA.

关 键 词：脲醛树脂-二氧化锆复合微球 亚氨基二乙酸 金属螫合亲和色谱 牛血清白蛋白(BSA) 

分 类 号：O652.63[理学—分析化学]