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Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology
Single B cell sampling following to direct gene amplification and transient expression in animal cells has been recognized as powerful monoclonal antibodies (mAbs) screening strategies. Here we report Ecobody technology which allows mAbs screening from single B cells in two days This technology uses...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6698955/ https://www.ncbi.nlm.nih.gov/pubmed/31544888 http://dx.doi.org/10.3390/antib7040038 |
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author | Ojima-Kato, Teruyo Morishita, Shiomi Uchida, Yoshino Nagai, Satomi Kojima, Takaaki Nakano, Hideo |
author_facet | Ojima-Kato, Teruyo Morishita, Shiomi Uchida, Yoshino Nagai, Satomi Kojima, Takaaki Nakano, Hideo |
author_sort | Ojima-Kato, Teruyo |
collection | PubMed |
description | Single B cell sampling following to direct gene amplification and transient expression in animal cells has been recognized as powerful monoclonal antibodies (mAbs) screening strategies. Here we report Ecobody technology which allows mAbs screening from single B cells in two days This technology uses Escherichia coli cell-free protein synthesis (CFPS) for mAb expression. In the CFPS step, we employed our original techniques: (1) ‘Zipbody’ as a modified Fab (fragment of antigen binding) format, in which the active Fab formation is facilitated by adhesive leucine zipper peptides fused at the C-termini of the light and heavy chains; and (2) an N-terminal SKIK peptide tag that can markedly increase protein production. By the Ecobody technology, we demonstrated rapid screening of antigen specific mAbs from immunized rabbits and Epstein-Barr Virus infected human B cells. We further obtained rabbit mAbs in E. coli expression system yielding to 8.5 mg of purified proteins from 1 L bacterial culture. |
format | Online Article Text |
id | pubmed-6698955 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-66989552019-09-05 Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology Ojima-Kato, Teruyo Morishita, Shiomi Uchida, Yoshino Nagai, Satomi Kojima, Takaaki Nakano, Hideo Antibodies (Basel) Article Single B cell sampling following to direct gene amplification and transient expression in animal cells has been recognized as powerful monoclonal antibodies (mAbs) screening strategies. Here we report Ecobody technology which allows mAbs screening from single B cells in two days This technology uses Escherichia coli cell-free protein synthesis (CFPS) for mAb expression. In the CFPS step, we employed our original techniques: (1) ‘Zipbody’ as a modified Fab (fragment of antigen binding) format, in which the active Fab formation is facilitated by adhesive leucine zipper peptides fused at the C-termini of the light and heavy chains; and (2) an N-terminal SKIK peptide tag that can markedly increase protein production. By the Ecobody technology, we demonstrated rapid screening of antigen specific mAbs from immunized rabbits and Epstein-Barr Virus infected human B cells. We further obtained rabbit mAbs in E. coli expression system yielding to 8.5 mg of purified proteins from 1 L bacterial culture. MDPI 2018-11-07 /pmc/articles/PMC6698955/ /pubmed/31544888 http://dx.doi.org/10.3390/antib7040038 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Ojima-Kato, Teruyo Morishita, Shiomi Uchida, Yoshino Nagai, Satomi Kojima, Takaaki Nakano, Hideo Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology |
title | Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology |
title_full | Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology |
title_fullStr | Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology |
title_full_unstemmed | Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology |
title_short | Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology |
title_sort | rapid generation of monoclonal antibodies from single b cells by ecobody technology |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6698955/ https://www.ncbi.nlm.nih.gov/pubmed/31544888 http://dx.doi.org/10.3390/antib7040038 |
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