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KMID : 0545120150250040547
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Journal of Microbiology and Biotechnology
2015 Volume.25 No. 4 p.547 ~ p.553
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Optimization of Yeast Surface-Displayed cDNA Library Screening for Low Abundance Targets
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Kim Ju-Hyung
Kim Hyung-Kyu
Jang Hye-Jeong
Kim Eun-Kyung
Kim Moon-Kyu
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Abstract
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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.
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KEYWORD
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cDNA library, yeast surface display, tumor-associated antigen, hepatocellular carcinoma
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