生物学杂志

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离子交换配基密度对蛋白质吸附行为的影响

  

  1.  
    1. 山东农业大学食品科学与工程学院;

    2. 中国科学院过程工程研究所生化工程国家重点实验室,

  • 出版日期:2016-04-18 发布日期:2016-04-18
  • 通讯作者: 王明林,教授,主要从事仪器分析方法研究,E-mail: mlwang@ sdau.edu. cn;张贵锋,研究员,主要从事生物大分子分离与分析方法研究,E-mail: gfzhang@ ipe.ac. cn
  • 作者简介:杨小雁,硕士研究生,主要从事界面表征方法研究,E-mail: inezyoung@163.com
  • 基金资助:
     
    国家青年科学基金( 21306205) ; 863 项目( 2014AA022109)

Effect of the ion exchange chromatography ligand density on the absorption behavior of proteins

  1.  
     1. College of Food Science and Engineering,Shandong Agriculture University,Taian 271018;

    2. National Key Laboratory of Biochemical Engineering,Institute of Process Engineering,Chinese Academy of Sciences,Beijing 100190,Chinav

  • Online:2016-04-18 Published:2016-04-18

摘要: 利用双偏振极化干涉测量仪(DPI)研究了离子交换配基密度对蛋白质吸附行为的影响。用N,N-二乙基-3-氨基丙基三甲氧基硅烷(DAPTMS)对DPI芯片进行修饰,利用X 射线光电子能谱表征了芯片上配基密度的不同。以溶菌酶、细胞色素C、糜蛋白酶、牛血清白蛋白和免疫球蛋白为模型蛋白,利用DPI研究了五种蛋白质在不同配基密度界面上吸附量、吸附密度和空间形态。结果表明,随着配基密度的增加,蛋白在界面上的吸附量、吸附密度随之增加,起始吸附速率随之增大;蛋白的空间形态与配基密度密切相关,特别是免疫球蛋白,蛋白层厚度远小于其分子直径,空间形态发生变化。证明配基密度是影响界面上蛋白质吸附量、吸附密度和空间形态等吸附行为的关键因素。

关键词: 离子交换色谱, 配基密度, 吸附行为, 双偏振极化干涉测量仪

Abstract:  The effect of ligand density on the adsorption behavior of proteins was investigated using dual polarization interferometry (DPI).The sensor chips of DPI were modified with 3-aminopropy-l-triethoxysilane (DAPTMS). The ligand densities on the modified chips were compared using X-ray photoelectron spectroscopy.Lysozyme, myoglobin, chymotrypsin, bovine serum albumin and immunoglobulin were used as model proteins. The adsorption behaviors of five proteins, such as adsorption amount, adsorption density and spatial structure, on the modified chips were characterized using DPI. It was found that with the increase of ligand density, the adsorption amount and density of proteins at the interface increase. Meanwhile, the initial adsorption rate grows. The spatial structure of proteins, especially immunoglobulin, was closely related with ligand density. The thickness of adsorption immunoglobulin layer was much smaller than the molecular diameter. The results indicated that the ligand density on the modified chip affected the amount, density and spatial structure of proteins on the interfaces.

Key words: ion exchange chromatography, ligand density, adsorption behavior, dual polarization polarization interferometer