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

doi:10.1016/j.jim.2020.112750

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

Department of Chemistry & Nanoscience

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

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 coli, mammalian cell lines, yeast, and plant. In addition, our RA-tag/47RA mAb combination showed high sensitivity and reliable affinity (K_D = 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 language	English
Article number	112750
Journal	Journal of Immunological Methods
Volume	479
DOIs	https://doi.org/10.1016/j.jim.2020.112750
State	Published - Apr 2020

Bibliographical note

Publisher Copyright:
© 2020 Elsevier B.V.

Keywords

Affinity purification
Epitope tag
Immunodetection
Monoclonal antibody

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10.1016/j.jim.2020.112750

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title = "Novel short peptide tag from a bacterial toxin for versatile applications",

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.",

keywords = "Affinity purification, Epitope tag, Immunodetection, Monoclonal antibody",

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

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year = "2020",

month = apr,

doi = "10.1016/j.jim.2020.112750",

language = "English",

volume = "479",

journal = "Journal of Immunological Methods",

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AU - Lee, Tae Hee

AU - Kim, Kwang Soo

AU - Kim, Jin Hee

AU - Jeong, Jae Ho

AU - Woo, Hye Ryun

AU - Park, So Ra

AU - Sohn, Myung Ho

AU - Lee, Hyeon Ju

AU - Rhee, Joon Haeng

AU - Cha, Sun Shin

AU - Hwang, Joo Hee

AU - Chung, Kyung Min

PY - 2020/4

Y1 - 2020/4

N2 - 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.

AB - 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.

KW - Affinity purification

KW - Epitope tag

KW - Immunodetection

KW - Monoclonal antibody

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DO - 10.1016/j.jim.2020.112750

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ER -

Novel short peptide tag from a bacterial toxin for versatile applications

Abstract

Bibliographical note

Keywords

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