高等学校化学学报

高等学校化学学报 ›› 2002, Vol. 23 ›› Issue (12): 2217.

• 研究论文 •    下一篇

具有甲状腺素脱碘酶活性的抗体酶的制备及性质研究

孙立伟, 刘立岩, 赵大庆   

  1. 中国科学院长春应用化学研究所稀土化学与物理开放实验室, 长春130022
  • 收稿日期:2002-03-26 出版日期:2002-12-24 发布日期:2002-12-24
  • 通讯作者: 赵大庆(1963年出生),男,博士,研究员,博士生导师,主要从事生物无机化学研究.
  • 基金资助:

    国家自然科学基金重大项目(批准号:29890280)资助

Preparation and Properties of Abzyme with Thyroxine Deiodinase

SUN Li-Wei, LIU Li-Yan, ZHAO Da-Qing   

  1. Key Laboratory of Rare Earth Chemistry and Physics, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China
  • Received:2002-03-26 Online:2002-12-24 Published:2002-12-24

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摘要： 首次以具有适当疏水性的甲状腺素衍生物O-methyl-T4为半抗原,用杂交瘤技术得到相应的抗体(6E8),又经过苯甲基磺酰氟和NaHSe的化学修饰引入了催化基团硒代半胱氨酸,制备出具有较高甲状腺素脱碘酶活力的抗体酶(Se-6E8).用SDS-PAGE电泳对Se-6E8的纯度进行了鉴定,同时研究了其解离常数、温度及pH等酶学性质.结果表明,Se-6E8活力为2010U/μmolprotein,纯度达到电泳纯,催化反应的最适温度为57℃,最适pH为82.

关键词: 硒, 甲状腺素, 脱碘酶, 抗体酶

Abstract: The abzyme (Se-6E8) with a higher thyroxine deiodinase activity was prepared by modifying the serine residues of monoclonal antibody (6E8) with phenylmethanesulfonyl fluoride and sodium hydrogen selenide, and the 6E8 against O-methyl-T4, which is a kind of thyroxine derivatives and was taken as a hapten for the first time. Two bands were found corresponding to the 5.5 kD heavy chain and the 2.7 kD light chain respectively by SDS-PAGE. The characteristics of dissociation constants, pH, and temperature were also studied. The results show that the activity of Se-6E8 is 2010 U/μmol protein , and the proper temperature and pH of the catalytic reactions is 57 ℃ and 8.2 respectively.

Key words: Selenium, Thyroxine, Deiodinase, Abzyme

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