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Novel short peptide tag from a bacterial toxin for versatile applications

Title
Novel short peptide tag from a bacterial toxin for versatile applications
Authors
Lee, Tae HeeKim, Kwang SooKim, Jin HeeJeong, Jae-HoWoo, Hye RyunPark, So RaSohn, Myung-HoLee, Hyeon JuRhee, Joon HaengCha, Sun-ShinHwang, Joo-HeeChung, Kyung Min
Ewha Authors
차선신
SCOPUS Author ID
차선신scopus
Issue Date
2020
Journal Title
JOURNAL OF IMMUNOLOGICAL METHODS
ISSN
0022-1759JCR Link

1872-7905JCR Link
Citation
JOURNAL OF IMMUNOLOGICAL METHODS vol. 479
Keywords
Epitope tagMonoclonal antibodyImmunodetectionAffinity purification
Publisher
ELSEVIER
Indexed
SCIE; SCOPUS WOS scopus
Document Type
Article
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/MARTX(Vv). 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 coll., mammalian cell lines, yeast, and plant. In addition, our RA-tag/47RA mAb combination showed high sensitivity and reliable affinity (K-D = 5.90 x 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.
DOI
10.1016/j.jim.2020.112750
Appears in Collections:
자연과학대학 > 화학·나노과학전공 > Journal papers
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