1. AviTrap: A novel solution to achieve complete biotinylation.
- Author
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Lu C, Bevers J, Tyagi T, To H, Lin M, Ti S, Nakamura G, Lin W, Chen Y, Wu Y, Li H, Wu J, and Wang F
- Subjects
- Recombinant Proteins chemistry, Recombinant Proteins isolation & purification, Recombinant Proteins metabolism, Humans, Biotin chemistry, Animals, Carbon-Nitrogen Ligases chemistry, Carbon-Nitrogen Ligases metabolism, Biotinylation, Antibodies, Monoclonal chemistry
- 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., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Lu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Published
- 2024
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