色谱

色谱

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高效液相色谱-基质辅助激光解吸电离串联飞行时间质谱分离鉴定牛乳铁蛋白素

安美忱, 刘宁*   

  1. 东北农业大学乳品科学教育部重点实验室, 黑龙江 哈尔滨 150030
  • 收稿日期:2009-06-30 修回日期:2009-08-30 出版日期:2010-02-28 发布日期:1981-06-25
  • 通讯作者: 刘宁
  • 基金资助:
    东北农业大学创新团队基金项目No. CXT007-3-1

Separation and identification of bovine lactoferricin by high performance liquid chromatography-matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry

AN Meichen, LIU Ning*   

  1. Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China
  • Received:2009-06-30 Revised:2009-08-30 Online:2010-02-28 Published:1981-06-25
  • Contact: LIU Ning

PDF

542

被引次数

9

摘要： 建立了高效液相色谱-基质辅助激光解吸电离串联飞行时间质谱(MALDI-TOF/TOF MS)分离鉴定牛乳铁蛋白素(bovine lactoferricin, LfcinB)的方法。采用胃蛋白酶酶解牛乳铁蛋白,酶解液离心后取上清液,经过离子交换色谱、反相液相色谱、透析等技术分离,对分离得到的目标产物进行抗菌活性分析、蛋白含量测定和MALDI-TOF/TOF MS鉴定。分离得到高活性的LfcinB,其相对分子质量为3124.89,蛋白含量为18.20 μg/mL。本方法具有精度高、分析速度快和分辨能力强等优点,是其他传统的分析鉴定LfcinB方法所无法比拟的,为进一步研究LfcinB奠定了基础。

关键词: 高效液相色谱, 基质辅助激光解吸电离串联飞行时间质谱, 牛乳铁蛋白 , 牛乳铁蛋白素

Abstract: A high performance liquid chromatography-matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry (HPLC-MALDI-TOF/TOF MS) method was developed for the separation and identification of bovine lactoferricin (LfcinB). Bovine lactoferrin was hydrolyzed by pepsin and then separated by ion exchange chromatography and reversed-phase liquid chromatography (RP-LC). The antibacterial activities of the fractions from RP-LC separation were determined and the protein concentration of the fraction with the highest activity was measured, whose sequence was indentified by MALDI-TOF/TOF MS. The relative molecular mass of LfcinB was 3124.89 and the protein concentration was 18.20 μg/mL. The method of producing LfcinB proposed in this study has fast speed, high accuracy and high resolution.

Key words: bovine lactoferricin, bovine lactoferrin , matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry (MALDI-TOF/TOF MS), high performance liquid chromatography (HPLC)