PDF(Pubmed)
Abstract：
Site specific biotinylation of AviTagged recombinant proteins using BirA enzyme is a widely used protein labeling technology. However, due to the incomplete biotinylation reactions and the lack of a purification method specific for the biotinylated proteins, it is challenging to purify the biotinylated sample when mixed with the non-biotinylated byproduct. Here, we have developed a monoclonal antibody that specifically recognizes the non-biotinylated AviTag but not the biotinylated sequence. After a ten-minute incubation with the resin that is conjugated with the antibody, the non-biotinylated AviTagged protein is trapped on the resin while the fully biotinylated material freely passes through. Therefore, our AviTrap (anti-AviTag antibody conjugated resin) provides an efficient solution for enriching biotinylated AviTagged proteins via a simple one-step purification.
摘要：
使用BirA酶对AviTagged重组蛋白进行位点特异性生物素化是一种广泛使用的蛋白质标记技术。然而,由于不完全的生物素化反应和缺乏特定于生物素化蛋白质的纯化方法，当与非生物素化的副产物混合时,纯化生物素化的样品是具有挑战性的。这里,我们已经开发了一种单克隆抗体，该抗体特异性识别非生物素化的AviTag，但不识别生物素化的序列。用与抗体结合的树脂孵育十分钟后，当完全生物素化的材料自由通过时,非生物素化的适合标记的蛋白质被捕获在树脂上。因此,我们的AviTrap（抗AviTag抗体缀合树脂）提供了通过简单的一步纯化富集生物素化AviTagged蛋白的有效解决方案。
