Yoshida Wataru, Sode Koji, Ikebukuro Kazunori
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Koganei, Tokyo, Japan.
Biotechnol Lett. 2008 Mar;30(3):421-5. doi: 10.1007/s10529-007-9575-3. Epub 2007 Oct 31.
We have developed an aptameric enzyme subunit (AES) for immunoglobulin E (IgE) sensing. AES is an artificial enzyme subunit constructed from two different aptamers and does not require any modification. Using the AES, the target molecule can be detected by measuring enzymatic activity in homogeneous solution. We connected IgE-binding aptamer and its complementary strand to split thrombin-inhibiting aptamer. The hybrid of these two oligonucleotides inhibited thrombin activity and it decreased in the presence of IgE. We were able to detect IgE by using this AES in homogeneous solution with a detection limit of 50 pmol.
我们开发了一种用于免疫球蛋白E(IgE)传感的适体酶亚基(AES)。AES是由两种不同适体构建而成的人工酶亚基,无需任何修饰。使用AES,可通过测量均相溶液中的酶活性来检测目标分子。我们将IgE结合适体及其互补链连接到裂解的凝血酶抑制适体上。这两种寡核苷酸的杂交体抑制了凝血酶活性,并且在IgE存在时其活性降低。我们能够在均相溶液中使用这种AES检测IgE,检测限为50皮摩尔。