Novel short peptide tag from a bacterial toxin for versatile applications
- 주제(키워드) Epitope tag , Monoclonal antibody , Immunodetection , Affinity purification
- 주제(기타) Biochemical Research Methods
- 주제(기타) Immunology
- 설명문(일반) [Lee, Tae Hee; Lee, Hyeon Ju; Chung, Kyung Min] Jeonbuk Natl Univ, Dept Microbiol & Immunol, Med Sch, Jeonju 54896, Jeonbuk, South Korea; [Lee, Tae Hee; Chung, Kyung Min] Jeonbuk Natl Univ, Inst Med Sci, Med Sch, Jeonju 54896, Jeonbuk, South Korea; [Kim, Kwang Soo; Jeong, Jae-Ho; Rhee, Joon Haeng] Chonnam Natl Univ, Dept Microbiol, Med Sch, Hwasun Gun 58128, South Korea; [Kim, Kwang Soo; Jeong, Jae-Ho; Rhee, Joon Haeng] Chonnam Natl Univ, Combinatorial Tumor Immunotherapy Med Res Ctr, Med Sch, Hwasun Gun 58128, South Korea; [Kim, Jin Hee] Jeju Natl Univ, Subtrop Hort Res Inst, Jeju 63243, South Korea; [Kim, Jin Hee; Woo, Hye Ryun] DGIST, Dept New Biol, Daegu 42988, South Korea; [Park, So Ra; Sohn, Myung-Ho] Osong Med Innovat Fdn, New Drug Dev Ctr, Cheongju 28160, Chungbuk, South Korea; [Rhee, Joon Haeng] Chonnam Natl Univ, Clin Vaccine R&D Ctr, Med Sch, Hwasun Gun 58128, South Korea; [Rhee, Joon Haeng] Vaxcell Bio Therapeut, Hwasun Gun 58141, South Korea; [Cha, Sun-Shin] Ewha Womans Univ, Dept Chem & Nanosci, Seoul 03760, South Korea; [Hwang, Joo-Hee] Jeonbuk Natl Univ, Dept Internal Med, Med Sch, Jeonju 54896, Jeonbuk, South Korea; [Hwang, Joo-Hee; Chung, Kyung Min] Jeonbuk Natl Univ, Res Inst Clin Med, Biomed Res Inst, Jeonbuk Natl Univ Hosp, Jeonju 54907, Jeonbuk, South Korea
- 등재 SCIE, SCOPUS
- 발행기관 ELSEVIER
- 발행년도 2020
- 총서유형 Journal
- URI http://www.dcollection.net/handler/ewha/000000169555
- 본문언어 영어
- Published As https://dx.doi.org/10.1016/j.jim.2020.112750
- PubMed https://pubmed.ncbi.nlm.nih.gov/31981564
초록/요약
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.
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