Optimization of yeast surface-displayed cDNA library screening for low abundance targets.
J Microbiol Biotechnol
; 25(4): 547-53, 2015 Apr.
Article
in En
| MEDLINE
| ID: mdl-25381740
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.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Gene Library
/
Two-Hybrid System Techniques
/
Cell Surface Display Techniques
Type of study:
Diagnostic_studies
/
Screening_studies
Limits:
Humans
Language:
En
Journal:
J Microbiol Biotechnol
Year:
2015
Document type:
Article