[Identification of protein antigen for antibody DBD02 by biopanning of T7 phage display library]
Xiang He1, Jin-ju Yang, Ying Liu
1Department of Immunology, Beijing Institute of Radiation Medicine, Beijing 100850, China. hexiang_spring@yahoo.com
Aim:
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).
Conclusion:
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.
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