Adv Pharm Bull. 2016;6(4):563-571 doi: 10.15171/apb.2016.070
Development of a Novel Human Single Chain Antibody Against EGFRVIII
Antigen by Phage Display Technology
Purpose: EGFRvIII as the most
common mutant variant of the epidermal growth factor receptor is resulting from
deletion of exons 2–7 in the coding sequence and junction of exons 1 and 8
through a novel glycine residue.
EGFRvIII is highly expressed in glioblastoma, carcinoma of the
breast, ovary, and lung but not in normal cells. The aim of the present
study was identification of a novel
single chain antibody against EGFRvIII as a promising target
for cancer therapy.
this study, a
synthetic peptide corresponding to EGFRvIII protein was used for screening a naive human scFv phage
library. A novel five-round selection strategy was used for enrichment of rare
Results: After five rounds of screening, six positive
scFv clones against EGFRvIII were selected using monoclonal phage ELISA, among
them, only three clones had expected size in PCR reaction. The specific
interaction of two of the scFv clones with EGFRvIII was confirmed by indirect
ELISA. One phage clone with higher affinity in scFv ELISA was purified for
further analysis. The purity of the produced scFv antibody was confirmed using
SDS-PAGE and Western blotting analyses.
Conclusion: In the present study, a human anti- EGFRvIII scFv with
high affinity was first identified from a scFv phage library. This study can be
the groundwork for developing more effective diagnostic and therapeutic agents
against EGFRvIII expressing cancers.