17th International Conference on Veterinary and Biomedical Sciences , 2015-10-26

Title : ( Application of phage display antibody library for Production of single-chain antibodies against common epitopes of ErbB1 and ErbB2 )

Authors: GHOLAMREZA HASHEMI TABAR , Reza Valadan , Rafiei. A , Mohammad Reza Bassami ,

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Abstract

Cancer cells use a complex multilayer network of epidermal growth factor receptors (EGFRs) signaling pathways to support their survival and growth. The overlapping networks of EGFRs signaling pathways account for the failure of most ErbB-targeted therapies. The aim of this study was to enrich a pool of recombinant antibody fragments against common epitopes of ErbB1 and ErbB2 in order to simultaneous blockade of ErbBs signaling pathways. ErbB1 and ErbB2 were expressed stably in VERO cells. Selection of recombinant antibodies was performed on live cells expressing either of ErbB1 and ErbB2 receptors using subtractive phage display approch. The results of PCR and DNA fingerprinting in the last round of panning showed that most clones contained insert (80% and 85% for ErbB1 and ErbB2 respectively) with an identical restriction pattern. The selected clones showed positive reaction to both ErbB1 and ErbB2 receptors in phage-ELISA test. Furthermore, the resulting soluble antibody fragments recognized common epitopes of both immunoprecipitated ErbB1 and ErbB2 in western blot. Additionally, the antibodies directed against the dimerization domain of ErbB1 demonstrated a significant absorbance in EGF-stimulated VERO/ErbB1 cells than non-stimulated cells (1.91 and 1.09 respectively). Moreover, the results of dimerization inhibition test showed that these antibodies blocked ErbB1 and ErbB2 dimerization on the surface of ErbB1 and ErbB2 expressing VERO cells. Regarding the importance of pan-ErbB approach to cancer therapy, the antibodies developed here might provide novel therapeutics for simultaneous blockade of ErbBs signaling pathways.

Keywords

, phage display antibody library, Single-chain antibody, ErbB1, ErbB2, Epitope
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@inproceedings{paperid:1053144,
author = {HASHEMI TABAR, GHOLAMREZA and Valadan, Reza and Rafiei. A and Bassami, Mohammad Reza},
title = {Application of phage display antibody library for Production of single-chain antibodies against common epitopes of ErbB1 and ErbB2},
booktitle = {17th International Conference on Veterinary and Biomedical Sciences},
year = {2015},
location = {بارسلونا},
keywords = {phage display antibody library; Single-chain antibody; ErbB1; ErbB2; Epitope},
}

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%0 Conference Proceedings
%T Application of phage display antibody library for Production of single-chain antibodies against common epitopes of ErbB1 and ErbB2
%A HASHEMI TABAR, GHOLAMREZA
%A Valadan, Reza
%A Rafiei. A
%A Bassami, Mohammad Reza
%J 17th International Conference on Veterinary and Biomedical Sciences
%D 2015

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