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http://dx.doi.org/10.4014/jmb.1409.09079

Optimization of Yeast Surface-Displayed cDNA Library Screening for Low Abundance Targets  

Kim, Juhyung (Department of Immunology, School of Medicine, Kyungpook National University)
Kim, Hyung Kyu (Department of Immunology, School of Medicine, Kyungpook National University)
Jang, Hye Jeong (Department of Immunology, School of Medicine, Kyungpook National University)
Kim, Eunkyung (Department of Immunology, School of Medicine, Kyungpook National University)
Kim, Moon Kyu (Department of Immunology, School of Medicine, Kyungpook National University)
Publication Information
Journal of Microbiology and Biotechnology / v.25, no.4, 2015 , pp. 547-553 More about this Journal
Abstract
The yeast surface-displayed cDNA library has been used to identify unknown antigens. However, when unknown target antigens show moderate-to-low abundance, some modifications are needed in the screening process. In this study, a directional random-primed cDNA library was used to increase the number of candidates for the unknown antigen. To avoid the loss of target yeast clones that express proteins at a low frequency in the cDNA library, a comprehensive monitoring system based on magnetic-activated cell sorting, fluorescence-activated cell sorting, and immunofluorescence was established, and a small number of target yeast cells was successfully enriched. These results showed that our optimized method has potential application for identifying rare unknown antigens of the human monoclonal antibody.
Keywords
cDNA library; yeast surface display; tumor-associated antigen; hepatocellular carcinoma;
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