Naqvi Azmi, Nahar Pradip
Institute of Genomics and Integrative Biology (formerly, Centre for Biochemical Technology), Delhi University Campus, Mall Road, Delhi 110007, India.
Anal Biochem. 2004 Apr 1;327(1):68-73. doi: 10.1016/j.ab.2003.11.026.
We report a rapid and versatile procedure for the preparation of photoreactive polymers and light-induced immobilization of proteins onto such polymers. Photoreactive controlled-pore glass, silica gel, glass slide, and polystyrene microtiter plate are prepared in 40-60s by microwave irradiation of the respective amino polymers and 1-fluoro-2-nitro-4-azidobenzene. Azido group, now part of the polymer, yields highly reactive nitrene under ultraviolet (UV) light at 365 nm. Thus, when photoreactive polymer and horseradish peroxidase or glucose oxidase are exposed to UV light, the reactive nitrene immobilizes the protein molecules in 10 to 20 min through covalent bonding. As nitrene has a property of inserting into C-H bond, the method may find potential applications for immobilization of biomolecules irrespective of their functional groups.
我们报道了一种制备光反应性聚合物以及将蛋白质光诱导固定到此类聚合物上的快速且通用的方法。通过对相应的氨基聚合物和1-氟-2-硝基-4-叠氮苯进行微波辐照，在40 - 60秒内即可制备出光反应性可控孔玻璃、硅胶、载玻片和聚苯乙烯微量滴定板。现在作为聚合物一部分的叠氮基团在365 nm紫外光下会产生高活性的氮烯。因此，当光反应性聚合物与辣根过氧化物酶或葡萄糖氧化酶暴露于紫外光时，活性氮烯会在10到20分钟内通过共价键固定蛋白质分子。由于氮烯具有插入C - H键的特性，该方法可能在固定生物分子方面具有潜在应用，而不论其官能团如何。
