AviTrap: A novel solution to achieve complete biotinylation

• Published in: PloS one Vol. 19; no. 4; p. e0297122
• Main Authors: Lu, Cheng, Bevers, Jack, Tyagi, Tulika, To, Hao, Lin, May, Ti, Shu, Nakamura, Gerry, Lin, WeiYu, Chen, Yongmei, Wu, Yan, Li, Hong, Wu, Jiansheng, Wang, Feng
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 25.04.2024; Public Library of Science (PLoS)
• Subjects: Analysis; Animals; Antibodies, Monoclonal - chemistry; Biology and Life Sciences; Biotin; Biotin - chemistry; Biotinylation; Carbon-Nitrogen Ligases - chemistry; Carbon-Nitrogen Ligases - metabolism; Chemical synthesis; Dosage and administration; Humans; Medicine and Health Sciences; Monoclonal antibodies; Physical Sciences; Recombinant proteins; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Research and Analysis Methods; United States
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0297122

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.