AviTrap: A novel solution to achieve complete biotinylation

Author:

Lu Cheng,Bevers JackORCID,Tyagi Tulika,To Hao,Lin May,Ti Shu,Nakamura Gerry,Lin WeiYu,Chen Yongmei,Wu Yan,Li Hong,Wu Jiansheng,Wang FengORCID

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.

Publisher

Public Library of Science (PLoS)

Reference30 articles.

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