TY - JOUR
T1 - Rapid antibody selection by mRNA display on a microfluidic chip
AU - Tabata, Noriko
AU - Sakuma, Yuko
AU - Honda, Yumiko
AU - Doi, Nobuhide
AU - Takashima, Hideaki
AU - Miyamoto-Sato, Etsuko
AU - Yanagawa, Hiroshi
N1 - Funding Information:
Grant-in-Aid for Scientific Research; Special Coordination Fund grant from the MEXT of Japan; the Industrial Technology Research Grant Program in ‘05 from the NEDO of Japan. Funding for open access charge: Keio University.
PY - 2009
Y1 - 2009
N2 - 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.
AB - 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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U2 - 10.1093/nar/gkp184
DO - 10.1093/nar/gkp184
M3 - Article
C2 - 19336414
AN - SCOPUS:65849248629
SN - 0305-1048
VL - 37
JO - Nucleic acids research
JF - Nucleic acids research
IS - 8
M1 - e64
ER -