Novel short peptide tag from a bacterial toxin for versatile applications
| Autor: | Hye Ryun Woo, Kyung Min Chung, Joo-Hee Hwang, Jae-Ho Jeong, Tae Hee Lee, Kwangsoo Kim, Sun Shin Cha, Hyeon Ju Lee, Jin Hee Kim, Myung-Ho Sohn, So Ra Park, Joon Haeng Rhee |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
medicine.drug_class Immunology Bacterial Toxins Antibody Affinity Peptide Enzyme-Linked Immunosorbent Assay medicine.disease_cause Monoclonal antibody Sensitivity and Specificity Epitope law.invention 03 medical and health sciences Epitopes 0302 clinical medicine Affinity chromatography Protein Domains law Protein purification medicine Escherichia coli Immunology and Allergy Animals Humans Peptide sequence Vibrio cholerae Vibrio vulnificus chemistry.chemical_classification Antibodies Monoclonal Recombinant Proteins 030104 developmental biology chemistry Biochemistry Recombinant DNA Peptides 030215 immunology |
| Zdroj: | Journal of immunological methods. 479 |
| ISSN: | 1872-7905 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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