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Nucleic Acids Research Advance Access published online on March 30, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp184
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© 2009
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Methods Online

Rapid antibody selection by mRNA display on a microfluidic chip

Noriko Tabata, Yuko Sakuma, Yumiko Honda, Nobuhide Doi, Hideaki Takashima, Etsuko Miyamoto-Sato and Hiroshi Yanagawa*

Department of Biosciences and Informatics, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama 223-8522, Japan

*To whom correspondence should be addressed. Tel: +81 45 566 1775; Fax: +81 45 566 1440; Email: hyana{at}bio.keio.ac.jp

Received January 13, 2009. Revised March 4, 2009. Accepted March 8, 2009.

In vitro antibody-display technologies are powerful approaches for isolating monoclonal antibodies from recombinant antibody libraries. However, these display techniques require several rounds of affinity selection which is time-consuming. Here, we combined mRNA display with a microfluidic system for in vitro selection and evolution of antibodies and achieved ultrahigh enrichment efficiency of 106- to 108-fold per round. After only one or two rounds of selection, antibodies with high affinity and specificity were obtained from naïve and randomized single-chain Fv libraries of ~1012 molecules. Furthermore, we confirmed that not only protein–protein (antigen–antibody) interactions, but also protein–DNA and protein–drug interactions were selected with ultrahigh efficiencies. This method will facilitate high-throughput preparation of antibodies and identification of protein interactions in proteomic and therapeutic fields.


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T. Sumida, N. Doi, and H. Yanagawa
Bicistronic DNA display for in vitro selection of Fab fragments
Nucleic Acids Res., September 29, 2009; (2009) gkp776v1.
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