Novel short peptide tag from a bacterial toxin for versatile applications

Tae Hee Lee, Kwang Soo Kim, Jin Hee Kim, Jae Ho Jeong, Hye Ryun Woo, So Ra Park, Myung Ho Sohn, Hyeon Ju Lee, Joon Haeng Rhee, Sun Shin Cha, Joo Hee Hwang, Kyung Min Chung

Research output: Contribution to journalArticlepeer-review

Abstract

The specific recognition between a monoclonal antibody (mAb) and its epitope can be used in a tag system that has proved valuable in a wide range of biological applications. Herein, we describe a novel tag called RA-tag that is composed of a seven amino acid sequence (DIDLSRI) and recognized by a highly specific mAb, 47RA, against the bacterial toxin Vibrio vulnificus RtxA1/MARTXVv. By using recombinant proteins with the RA-tag at the N-terminal, C-terminal, or an internal site, we demonstrated that the tag system could be an excellent biological system for both protein purification and protein detection in enzyme-linked immunosorbent, Western blot, flow cytometry, and immunofluorescence staining analyses in Escherichia coli, mammalian cell lines, yeast, and plant. In addition, our RA-tag/47RA mAb combination showed high sensitivity and reliable affinity (KD = 5.90 × 10−8 M) when compared with conventional tags. Overall, our results suggest that the RA-tag system could facilitate the development of a broadly applicable tag system for biological research.

Original languageEnglish
Article number112750
JournalJournal of Immunological Methods
Volume479
DOIs
StatePublished - Apr 2020

Keywords

  • Affinity purification
  • Epitope tag
  • Immunodetection
  • Monoclonal antibody

Fingerprint

Dive into the research topics of 'Novel short peptide tag from a bacterial toxin for versatile applications'. Together they form a unique fingerprint.

Cite this