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Antibodies 2018, 7(4), 38; https://doi.org/10.3390/antib7040038

Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology

1
iBody Inc., Furo-cho 1, Chikusa-ku, Nagoya 464-0814, Japan
2
Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan
There authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 25 September 2018 / Revised: 25 October 2018 / Accepted: 31 October 2018 / Published: 7 November 2018
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Abstract

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. View Full-Text
Keywords: single B cell technology; rabbit antibodies; human antibodies single B cell technology; rabbit antibodies; human antibodies
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Ojima-Kato, T.; Morishita, S.; Uchida, Y.; Nagai, S.; Kojima, T.; Nakano, H. Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology. Antibodies 2018, 7, 38.

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