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化学学报
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化学学报 ›› 2008, Vol. 66 ›› Issue (13): 1572-1576. 上一篇    下一篇

研究论文

免疫亲和质谱法研究β2-微球蛋白抗原表位

万翠红 黎 根 刘志强 刘淑莹*   

  1. (中国科学院长春应用化学研究所 长春质谱中心 长春 130022)
  • 投稿日期:2008-01-06 修回日期:2008-03-01 发布日期:2008-07-14
  • 通讯作者: 刘淑莹

Characterization Epitope of the β2-Microglobulin with Immunoaffinity Mass Spectrometry

WAN, Cui-Hong LI, Gen LIU, Zhi-Qiang LIU, Shu-Ying*   

  1. (Changchun Center of Mass Spectrometry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences,
    Changchun 130022)
  • Received:2008-01-06 Revised:2008-03-01 Published:2008-07-14
  • Contact: LIU, Shu-Ying

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采用免疫亲和分离与质谱分析相结合的方法, 对β2-微球蛋白抗原表位进行了系统研究. 完整的抗原分子和已固定在载体(CNBr-activated Sepharose beads)上的单克隆抗体发生免疫亲和反应后, 用Endoproteinase Glu-C, Trypsin, Aminopeptidase M和carboxypeptidase Y四种不同的蛋白酶依次酶解抗原分子, 并采用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)技术对与抗体连接受保护而未发生水解的肽段进行了研究. 结果表明: β2-微球蛋白抗原表位位于整个蛋白分子氨基酸序列的61～67位, 即为SFYLLYY. 通过合成肽段的分析, 证明了SFYLLYY即为抗原表位, 与亲和质谱方法分析结果一致.

关键词: β2-微球蛋白, 抗原表位, 免疫亲和, 质谱

The approach for epitope mapping is the application of immunoaffinity separation and matrix assisted laser desorption/ionization (MALDI) mass spectrometry (MS). The epitope of β2-microglobulin was identified with this approach. The antigen β2-microglobulin was coupled to a monoclonal antibody, which was covalently immobilized on a cyanogen bromide (CNBr)-activated Sepharose bead. Consecutive digestion of the antigen with endoproteinase Glu-C, trypsin, aminopeptidase M and carboxypeptidase Y resulted in affinity-bound peptides containing epitope. The bound peptides were identified by MALDI-TOF MS. The epitope recognized by the monoclonal antibody was identified to be the peptide fragment 61～67 with the sequence SFYLLYY. Synthesized peptide indicated the same result.

Key words: β2-microglobulin, epitope, immunoaffinity, mass spectrometry