PLoS ONE (Jan 2024)

AviTrap: A novel solution to achieve complete biotinylation.

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

DOI
https://doi.org/10.1371/journal.pone.0297122
Journal volume & issue
Vol. 19, no. 4
p. e0297122

Abstract

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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.