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

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

Teruyo Ojima-Kato, Shiomi Morishita, Yoshino Uchida, Satomi Nagai, Takaaki Kojima, Hideo Nakano

Antibodies (Basel). 2018 Nov 7;7(4):38.

PMID: 31544888

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR42417579	Anti-Sprouty 2 (N-Terminal) from rabbit			Price

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