학술논문

AviTrap: A novel solution to achieve complete biotinylation.
Document Type
Article
Source
PLoS ONE. 4/25/2024, Vol. 19 Issue 4, p1-17. 17p.
Subject
*RECOMBINANT proteins
*MONOCLONAL antibodies
*ENZYMES
Language
ISSN
1932-6203
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. [ABSTRACT FROM AUTHOR]