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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Tabriz University of Medical Sciences
2016-12-01
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| Series: | Advanced Pharmaceutical Bulletin |
| Subjects: | |
| Online Access: | http://journals.tbzmed.ac.ir/APB/Manuscript/APB-6-563.pdf |
| _version_ | 1811281977777586176 |
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| author | Leila Rahbarnia Safar Farajnia Hossein Babaei Jafar Majidi Bahman Akbari Shiva Ahdi khosroshahi |
| author_facet | Leila Rahbarnia Safar Farajnia Hossein Babaei Jafar Majidi Bahman Akbari Shiva Ahdi khosroshahi |
| author_sort | Leila Rahbarnia |
| collection | DOAJ |
| description | 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.
Methods: In
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
specific clones.
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. |
| first_indexed | 2024-04-13T01:43:22Z |
| format | Article |
| id | doaj.art-b108709202224c4d88a17160f9105d5a |
| institution | Directory Open Access Journal |
| issn | 2228-5881 2251-7308 |
| language | English |
| last_indexed | 2024-04-13T01:43:22Z |
| publishDate | 2016-12-01 |
| publisher | Tabriz University of Medical Sciences |
| record_format | Article |
| series | Advanced Pharmaceutical Bulletin |
| spelling | doaj.art-b108709202224c4d88a17160f9105d5a2022-12-22T03:08:07ZengTabriz University of Medical SciencesAdvanced Pharmaceutical Bulletin2228-58812251-73082016-12-016456357110.15171/apb.2016.070APB_1354_20160718140917Development of a Novel Human Single Chain Antibody Against EGFRVIII Antigen by Phage Display TechnologyLeila Rahbarnia0Safar Farajnia1Hossein Babaei2Jafar Majidi3Bahman Akbari4Shiva Ahdi khosroshahi5Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.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. Methods: In 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 specific clones. 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.http://journals.tbzmed.ac.ir/APB/Manuscript/APB-6-563.pdfHuman single chain antibodyCancerEGFRvIIIPhage display |
| spellingShingle | Leila Rahbarnia Safar Farajnia Hossein Babaei Jafar Majidi Bahman Akbari Shiva Ahdi khosroshahi Development of a Novel Human Single Chain Antibody Against EGFRVIII Antigen by Phage Display Technology Advanced Pharmaceutical Bulletin Human single chain antibody Cancer EGFRvIII Phage display |
| title | Development of a Novel Human Single Chain Antibody Against EGFRVIII
Antigen by Phage Display Technology |
| title_full | Development of a Novel Human Single Chain Antibody Against EGFRVIII
Antigen by Phage Display Technology |
| title_fullStr | Development of a Novel Human Single Chain Antibody Against EGFRVIII
Antigen by Phage Display Technology |
| title_full_unstemmed | Development of a Novel Human Single Chain Antibody Against EGFRVIII
Antigen by Phage Display Technology |
| title_short | Development of a Novel Human Single Chain Antibody Against EGFRVIII
Antigen by Phage Display Technology |
| title_sort | development of a novel human single chain antibody against egfrviii antigen by phage display technology |
| topic | Human single chain antibody Cancer EGFRvIII Phage display |
| url | http://journals.tbzmed.ac.ir/APB/Manuscript/APB-6-563.pdf |
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