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 o...

Full description

Saved in:
Bibliographic Details
Published inJournal of microbiology and biotechnology Vol. 25; no. 4; pp. 547 - 553
Main Authors Kim, Juhyung, Kim, Hyung Kyu, Jang, Hye Jeong, Kim, Eunkyung, Kim, Moon Kyu
Format Journal Article
LanguageKorean
Published 한국미생물생명공학회 30.04.2015
Subjects
cDNA library
hepatocellular carcinoma
tumor-associated antigen
yeast surface display
tumor-associated antigen
yeast surface display
hepatocellular carcinoma
cDNA library
Online AccessGet full text

Cover

More Information
Summary: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.
Bibliography:The Korean Society for Applied Microbiology
KISTI1.1003/JNL.JAKO201523050399527
ISSN:1017-7825
1738-8872