Jahanshahi-Anbuhi Sana, Kannan Balamurali, Leung Vincent, Pennings Kevin, Liu Meng, Carrasquilla Carmen, White Dawn, Li Yingfu, Pelton Robert H, Brennan John D, Filipe Carlos D M
Biointerfaces Institute , McMaster University , 1280 Main St W , Hamilton , ON L8S 4L8 , Canada . Email:
Department of Chemical Engineering , McMaster University , 1280 Main St W , Hamilton , ON L8S 4L7 , Canada . Email:
Chem Sci. 2016 Mar 1;7(3):2342-2346. doi: 10.1039/c5sc04184h. Epub 2016 Jan 4.
Many biodetection systems employ labile enzymes and substrates that need special care, making it hard to routinely use them for point-of-care or field applications. In this work we provide a simple solution to this challenging problem through the creation of all-inclusive pullulan assay tablets. The proposed tablet system not only enhances the long-term stability of both enzymes and organic substrates, but also simplifies the assay procedure. The enhanced stability is attributed to two factors: the restriction of the molecular motion of proteins and impermeability to molecular oxygen afforded by the tables. These tablets dissolve rapidly upon addition to testing samples, making the test very easy to perform. Using the ATP-detecting luciferase-luciferin system as an example, we show that the tablet-based assay can achieve highly sensitive detection of ATP in biological samples and that the activity of the assay tablets remains unchanged for over a month at room temperature.
许多生物检测系统使用需要特殊处理的不稳定酶和底物，这使得它们难以常规用于即时检测或现场应用。在这项工作中，我们通过创建全包式支链淀粉检测片剂，为这个具有挑战性的问题提供了一个简单的解决方案。所提出的片剂系统不仅提高了酶和有机底物的长期稳定性，还简化了检测程序。稳定性的提高归因于两个因素：片剂对蛋白质分子运动的限制以及对分子氧的不渗透性。这些片剂在加入测试样品后迅速溶解，使测试非常易于执行。以检测ATP的荧光素酶-荧光素系统为例，我们表明基于片剂的检测可以在生物样品中实现对ATP的高灵敏度检测，并且检测片剂的活性在室温下一个多月内保持不变。
