Reference: Kim JY, et al. (2015) Optimization of yeast surface-displayed cDNA library screening for low abundance targets. J Microbiol Biotechnol 25(4):547-53

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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.

Reference Type
Journal Article | Research Support, Non-U.S. Gov't
Authors
Kim JY, Kim HK, Jang HJ, Kim EK, Kim MK
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