[Identification of protein antigen for antibody DBD02 by biopanning of T7 phage display library].
OBJECTIVE
To identify the specificity of mAb DBD02, we developed and optimized a new method by biopanning of T7 Select human liver cDNA phage display library.


METHODS
After two rounds of biopanning, eluted phages were subjected to Dot blot using mAb DBD02 as primary antibody. The positive PFUs (plaque forming unit) which recognized by DBD02 were further confirmed by Western blot, PCR and sequencing.


RESULTS
The antigen recognized by DBD02 was identified as alcohol dehydrogenase. And the epitope for mAb DBD02 was further mapped within a peptide of 22 amino acids (QDYKKPIQEVLKEMTDG-GVDFS).


CONCLUSIONS
Our data indicate that biopanning of T7 phage display library by mAbs is time, cost, and labor-saving and specific tool for protein antigen identification.
He
Xiang
X
Department of Immunology, Beijing Institute of Radiation Medicine, Beijing 100850, China. hexiang_spring@yahoo.com
Yang
Jin-ju
JJ
Liu
Ying
Y
Liu
Xiao-lan
XL
Chen
Yong
Y
Gao
Jian-en
JE
Sun
Qi-hong
QH
D000426:Alcohol Dehydrogenase / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000915:Antibody Affinity; D000918:Antibody Specificity; D000941:Antigens / Q000032:analysis* / Q000737:chemistry / Q000276:immunology*; D017123:Bacteriophage T7; D015153:Blotting, Western; D015723:Gene Library; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D013997:Time Factors