色谱

色谱 ›› 2014, Vol. 32 ›› Issue (10): 1029-1033.DOI: 10.3724/SP.J.1123.2014.06029

• 研究快报 • 上一篇    下一篇

可逆加成-断裂链转移策略制备可识别转铁蛋白的抗原决定基印迹微球

李沁然1,2, 杨开广1, 李森武1,2, 刘键熙1,2, 张丽华1, 梁振1, 张玉奎1   

  1. 1. 国家色谱研究分析中心, 分离分析化学重点实验室, 中国科学院大连化学物理研究所, 辽宁 大连 116023;
    2. 中国科学院大学, 北京 100039
  • 收稿日期:2014-06-21 出版日期:2014-10-08 发布日期:2014-09-29
  • 通讯作者: 张丽华，E-mail：lihuazhang@dicp.ac.cn.
  • 基金资助:

    国家自然科学基金项目（21375128）；国家高科技研究计划（“863”计划）项目（2012AA020202）.

Preparation of epitope imprinted particles for transferrin recognition by reversible addition fragmentation chain transfer strategy

LI Qinran1,2, YANG Kaiguang1, LI Senwu1,2, LIU Jianxi1,2, ZHANG Lihua1, LIANG Zhen1, ZHANG Yukui1   

  1. 1. National Chromatographic R. & A. Center, Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China;
    2. University of Chinese Academy of Sciences, Beijing 100039, China
  • Received:2014-06-21 Online:2014-10-08 Published:2014-09-29

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195

被引次数

11

摘要：

应用可逆加成-断裂链转移（RAFT）策略制备了一种抗原决定基表面印迹微球。这一工作以转铁蛋白的抗原决定基N端九肽作为模板，通过共价键合的方式固载于修饰了戊二醛的硅胶颗粒表面。然后以甲基丙烯酸、甲基丙烯酰羟乙酯为功能单体，甲叉基双丙烯酰胺为交联剂，偶氮二异丁腈（AIBN）为引发剂，N，N-二甲基甲酰胺为溶剂，在三硫酯试剂2-（十二烷基三硫代碳酸酯基）-2-甲基丙酸的调控下，于70 ℃进行活性-可控的聚合反应，制备得到分子印迹微球。该材料对模板抗原决定基的识别容量为2.36 mg/g，印迹因子为1.89；对转铁蛋白的识别容量为4.98 mg/g，印迹因子为1.61，120 min内可达到吸附平衡；在多蛋白质竞争识别中，该材料对转铁蛋白识别的印迹因子远高于细胞色素C、乳球蛋白等其他竞争蛋白质的印迹因子。以上结果证明，通过RAFT策略制备得到的抗原决定基分子印迹材料在对抗原决定基具有良好的识别能力的同时，对模板抗原决定基对应的转铁蛋白也具有优良的选择性、较高的识别容量和较快的识别速度。

关键词: 蛋白质, 分子印迹, 抗原决定基, 可逆加成-断裂链转移（RAFT）

Abstract:

A kind of novel epitope surface imprinted particles was prepared by the reversible addition fragmentation chain transfer (RAFT) strategy. The epitope of transferrin, N-terminal peptide of the protein with nine amino acid residues, was chosen as the template and immobilized with covalent interaction on the surface of silica particles through the truss arm glutaraldehyde. The living/controlled polymerization was initialed by 2,2'-azobisisobutyronitrile (AIBN) at 70 ℃ in the solution of N,N-dimethylformamide, with the regulation by triothioester agent 2-(dodecylthiocarbonothioylthio)-2-methylpropanoic acid. Methacrylic acid and 2-hydroxyethyl methacrylate were chosen as the functional monomers and N,N-methylenebisacrylamide was chosen as the cross-linker in this polymerization. For this material, the binding capacity of the nine residue peptide could reach 2.36 mg/g with the imprinting factor (IF) of 1.89, while that for transferrin could reach 4.98 mg/g with IF of 1.61. The equilibrium could be achieved in 120 min for the transferrin recognition. In multi-protein competitive recognition, the imprinted factor of transferrin was the highest in the mixture of transferrin and other competitive proteins, such as cytochrome C and β-lactoglobulin. The results indicated that these epitope surface imprinted particles with RAFT strategy could recognize not only the nine residue peptide but also the transferrin with good selectivity, high binding capacity and fast mass transfer.

Key words: epitope, molecularly imprinting, protein, reversible addition fragmentation chain transfer (RAFT)

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