生物素化
生物素
重组DNA
单克隆抗体
链霉亲和素
化学
色谱法
抗体
生物化学
分子生物学
生物
免疫学
基因
作者
Cheng Lü,Cheng Lü,Jack Bevers,Tulika Tyagi,Hodges To,Millicent Lin,Millicent Lin,Shu Ti,Grant Nakamura,Wei‐Yu Lin,Yongmei Chen,Yan Wu,Hong Li,Hong Li,Jiansheng Wu,Feng Wang,Feng Wang
出处
期刊:PLOS ONE
[Public Library of Science]
日期:2024-04-25
卷期号:19 (4): e0297122-e0297122
标识
DOI:10.1371/journal.pone.0297122
摘要
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.
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