Optimization of yeast surface-displayed cDNA library screening for low abundance targets.
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.
Department of Immunology, School of Medicine, Kyungpook National University, Daegu 700-422, Republic of Korea.
Juh-Yung
JY
Kim
Department of Immunology, School of Medicine, Kyungpook National University, Daegu 700-422, Republic of Korea.
Hyung Kyu
HK
Kim
Department of Immunology, School of Medicine, Kyungpook National University, Daegu 700-422, Republic of Korea.
Hye Jeong
HJ
Jang
Department of Immunology, School of Medicine, Kyungpook National University, Daegu 700-422, Republic of Korea.
Eun-Kyung
EK
Kim
Department of Immunology, School of Medicine, Kyungpook National University, Daegu 700-422, Republic of Korea.
Moon Kyu
MK
Kim
D045744:Cell Line, Tumor; D061505:Cell Surface Display Techniques; D005434:Flow Cytometry / Q000379:methods; D015723:Gene Library; D006801:Humans; D017422:Sequence Analysis, DNA; D020798:Two-Hybrid System Techniques