Institute of Genomics and Integrative Biology (CSIR), Mall Road, Delhi 110 007, India.
J Colloid Interface Sci. 2010 Feb 1;342(1):202-4. doi: 10.1016/j.jcis.2009.10.038. Epub 2009 Oct 22.
Herein, we demonstrate a simple, versatile and efficient method for immobilization of unmodified carbohydrates onto a solid surface. The method employs a photoreactive-cellulose membrane which is prepared by the introduction of a photoreactive group to a cellulose membrane through 1-fluoro-2-nitro-4-azidobenzene (FNAB). Upon exposure to UV light photo-reactive azido group of the activated cellulose membrane transforms itself into highly reactive nitrene which covalently binds to underivatized carbohydrates through an insertion reaction. Maximum immobilization of carbohydrate is achieved at the UV exposure time of 60min and carbohydrate concentration of 100microg/disk. The immobilized carbohydrate is assayed by affinity binding of biotin-labeled lectins. The amount of Con A required for detecting immobilized carbohydrate ranges from 4-10microg/ml. The binding affinities of the lectin to the immobilized carbohydrates are analyzed by calculating their IC(50) values. Overall, the present work demonstrates an efficient immobilization of underivatized carbohydrate onto cellulose surface and has the potential to be applied to other surfaces.
在此，我们展示了一种简单、通用且高效的方法，可将未修饰的碳水化合物固定到固体表面上。该方法采用了一种光反应性纤维素膜，通过将光反应性基团通过 1-氟-2-硝基-4-叠氮苯（FNAB）引入纤维素膜来制备。在暴露于紫外光下时，活化纤维素膜的光反应性叠氮基团会转化为高反应性氮烯，通过插入反应与未衍生的碳水化合物共价结合。在 UV 暴露时间为 60 分钟和碳水化合物浓度为 100μg/盘时，可实现最大的碳水化合物固定化。通过生物素标记的凝集素的亲和结合来测定固定化的碳水化合物。检测固定化碳水化合物所需的 Con A 量范围为 4-10μg/ml。通过计算它们的 IC（50）值来分析凝集素与固定化碳水化合物的结合亲和力。总的来说，本工作展示了一种高效的将未衍生的碳水化合物固定到纤维素表面的方法，并且有可能应用于其他表面。
