高等学校化学学报

高等学校化学学报 ›› 2005, Vol. 26 ›› Issue (5): 793.

• 研究论文 •    下一篇

离子交换介孔分子筛DEAE-SBA-15的制备及其用于分离纯化粉尘螨主要变应原Derf2

雷杰1, 张鲁雁2, 丁声颂2, 江世益2, 余承忠1, 赵东元1   

  1. 1. 复旦大学分子催化与创新材料实验室;
    2. 复旦大学化学系, 上海200433
  • 收稿日期:2004-12-14 出版日期:2005-05-10 发布日期:2005-05-10
  • 通讯作者: 赵东元(1963年出生),男,博士,教授,博士生导师,主要从事介孔分子筛的合成及应用的研究.E-mail:dyzhao@fudan.edu.cn E-mail:dyzhao@fudan.edu.cn
  • 基金资助:

    国家自然科学基金(批准号:20301004);上海华谊集团、上海市科委、上海市纳米中心基金资助.

Preparation of Ion Exchange Mesoporous Sieve DEAE-SBA-15 and Its Application to Separation and Purification of Der f 2 Allergen in Dermatophagoides Farinae

LEI Jie1, ZHANG Lu-Yan2, DING Sheng-Song2, JIANG Shi-Yi2, YU Cheng-Zhong1, ZHAO Dong-Yuan1   

  1. 1. Molecular Catalysis and Innovative Materials Lab;
    2. Department of Chemistry, Fudan University, Shanghai 200433, China
  • Received:2004-12-14 Online:2005-05-10 Published:2005-05-10

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被引次数

20

摘要： 通过3-含氧缩水甘油基-丙基-三甲氧基硅烷(GPTS)将二乙胺基乙基(DEAE)嫁接在介孔分子筛SBA-15的表面,制得弱阴离子交换层析剂DEAE-SBA-15,使其既具有分子筛作用,又具有离子交换作用的特点.经DEAE-SBA-15层析后,从粉尘螨代谢培养基中获得单一的洗脱峰,SDS-PAGE显示一条谱带,其相对分子质量为14000,表明经分离纯化所得到的蛋白质是粉尘螨主要变应原Derf2.对螨性哮喘患者皮肤点刺试验表明,所得Derf2仍具有变应原活性,且仅通过一次柱色谱即得Derf2,表明DEAE-SBA-15是一种较好的离子交换层析剂.

关键词: DEAE-SBA-15离子交换介孔分子筛, 固定相制备, 粉尘螨, 纯化, Derf2

Abstract: Mesoporous silica SBA-15 was functionalized by 2-diethylamino-ethanol(DEAE) through the (epoxide) ring opening of 3-glycidyloxy-propyl-trimethoxy silane grafted on the silica surface(denoted as DEAE-SBA-15). Subsequently, DEAE-SBA-15 was used as the weak anion exchange matrix for chromatography to purify (Der f 2) allergen from the spent culture medium extract of Dermatophagoides farinae, one of House dust mites that can cause heavy atopic diseases such as asthma and dermatitis. The structure of DEAE-SBA-15 was confirmed by 13C and 29Si CP MAS NMR and other methods. By SDS-PAGE, Der f 2 was estimated to have a molecular weight of 14 000 and appeared as the only separated protein band. The assay results of skin prick test showed that the allergic activity of Der f 2 was retained. The above results indicate that DEAE-SBA-15 materials are promising fixed-matrix candidates in liquid chromatography to purify proteins.

Key words: DEAE-SBA-15, Stationary phase preparation, Dermatophagoides farinae, Purification, Der f 2 allergen

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