Isfahan University of Medical Sciences

Science Communicator Platform

Stay connected! Follow us on X network (Twitter):
Share this content! On (X network) By
Improved Biological Activity of a Single Chain Antibody Fragment Against Human Epidermal Growth Factor Receptor 2 (Her2) Expressed in the Periplasm of Escherichia Coli Publisher Pubmed



Akbari V1 ; Sadeghi HMM1 ; Jafariandehkordi A1 ; Abedi D1, 2 ; Chou CP2
Authors
Show Affiliations
Authors Affiliations
  1. 1. Department of Pharmaceutical Biotechnology, Isfahan Pharmaceutical Research Center, Faculty of Pharmacy, Isfahan University of Medical Sciences, Hezar Jarib Avenue, Isfahan, Iran
  2. 2. Department of Chemical Engineering, University of Waterloo, 200 University Avenue, Waterloo, N2L 3G1, ON, Canada

Source: Protein Expression and Purification Published:2015


Abstract

A novel monoclonal antibody against human epidermal growth factor receptor 2 (HER2), i.e., pertuzumab (Perjeta®) developed by Genentech, has been verified to be effective in treating metastatic HER2-overexpressing breast cancer. The fact that the presence of the Fc region of the anti-HER2 is uncritical for growth inhibition of tumor cells suggests the potential biological activity of the associated antibody fragments. In the present study, we report functional expression of anti-HER2his-scFv, a single-chain variable fragment (scFv) derived from pertuzumab, in the periplasm of Escherichia coli and its purification. Biological activity of the soluble scFv produced in this manner was characterized using immunofluorescent staining, immunocytochemistry, flow cytometry and cytotoxicity assay. The effect of anti-HER2his-scFv on HER2 dimerization was also assessed by tyrosine kinase assay. It was observed that the purified scFv had a high specificity and affinity to HER2 receptors expressed on the surface of tumor cells with a selective cytotoxic effect on HER2-overexpressing SK-OV-3 cells. In addition, anti-HER2his-scFv was able to suppress phosphorylation of HER2 in the presence of heregulin. The results suggest that anti-HER2his-scFv can be a potential candidate for various therapeutic and diagnosis applications. © 2015 Elsevier Inc.
Related Docs
View other Related Docs
1. Functional Expression of a Single-Chain Antibody Fragment Against Human Epidermal Growth Factor Receptor 2 (Her2) in Escherichia Coli, Journal of Industrial Microbiology and Biotechnology (2014)
2. New Insights Into Affinity Proteins for Her2-Targeted Therapy: Beyond Trastuzumab, Biochimica et Biophysica Acta - Reviews on Cancer (2020)
3. Optimization of a Single-Chain Antibody Fragment Overexpression in Escherichia Coli Using Response Surface Methodology, Research in Pharmaceutical Sciences (2015)
4. Design and Construction of Scfv-Pe35kdel As a Novel Immunotoxin Against Human Epidermal Growth Factor Receptor 2 for Cancer Therapy, Biotechnology Letters (2023)
5. Production and Characterization of New Anti-Her2 Monoclonal Antibodies, Monoclonal Antibodies in Immunodiagnosis and Immunotherapy (2015)
6. Improvement of Solubility and Refolding of an Anti-Human Epidermal Growth Factor Receptor 2 Single-Chain Antibody Fragment Inclusion Bodies, Research in Pharmaceutical Sciences (2018)
7. Linear and Conformational B Cell Epitope Prediction of the Her 2 Ecd-Subdomain Iii by in Silico Methods, Asian Pacific Journal of Cancer Prevention (2012)
8. The Prognostic Significance of Her2-Neu Over Expression in Gastric Carcinomas, Iranian Journal of Cancer Prevention (2011)
Experts (# of related papers)
View all Related Experts
Vajihe Akbari (11)
Abbas Jafarian (5)
Other Related Docs
9. Construction and Periplasmic Expression of a Bispecific Tandem Scfv for Dual Targeting of Immune Checkpoints, Advanced Biomedical Research (2023)
10. Integrated Bioprocessing and Genetic Strategies to Enhance Soluble Expression of Anti-Her2 Immunotoxin in E. Coli, AMB Express (2024)
11. Clinical Applications of Cell Encapsulation Technology in Cell and Drug Delivery, Journal of Isfahan Medical School (2016)
12. Comparison of the Cytoplasmic and Periplasmic Production of Reteplase in Escherichia Coli, Preparative Biochemistry and Biotechnology (2013)
13. Blockade of Exosome Release Alters Her2 Trafficking to the Plasma Membrane and Gives a Boost to Trastuzumab, Clinical and Translational Oncology (2023)
14. Comparing Efficacy and Safety of P013, a Proposed Pertuzumab Biosimilar, With the Reference Product in Her2-Positive Breast Cancer Patients: A Randomized, Phase Iii, Equivalency Clinical Trial, BMC Cancer (2022)
15. Optimization of Buffer Additives for Efficient Recovery of Hgm-Csf From Inclusion Bodies Using Response Surface Methodology, Iranian Journal of Pharmaceutical Research (2020)
16. In Silico Design of Discontinuous Peptides Representative of B and T-Cell Epitopes From Her2-Ecd As Potential Novel Cancer Peptide Vaccines, Asian Pacific Journal of Cancer Prevention (2013)
17. Investigation of Her-2 Expression and Its Correlation With Clinicopathological Parameters and Overall Survival of Esophageal Squamous Cell Carcinoma Patients, Iranian Journal of Pathology (2020)
18. Improvement of Soluble Expression of Gm-Csf in the Cytoplasm of Escherichia Coli Using Chemical and Molecular Chaperones, Protein Expression and Purification (2019)
19. Trastuzumab-Conjugated Nanoparticles Composed of Poly(Butylene Adipate-Co-Butylene Terephthalate) Prepared by Electrospraying Technique for Targeted Delivery of Docetaxel, IET Nanobiotechnology (2019)
20. Evaluation of Immunohistochemistry Technique in Detecting Her2 Overexpression in Invasive Ductal Breast Carcinoma Using Fluorescence In-Situ Hybridization Method, Journal of Isfahan Medical School (2016)
21. Evaluation of Soluble Expression of Recombinant Granulocyte Macrophage Stimulating Factor (Rgm-Csf) by Three Different E. Coli Strains, Research in Pharmaceutical Sciences (2020)
22. Validity of Immunohistochemistry Method in Predicting Her-2 Gene Status and Association of Clinicopathological Variables With It in Invasive Breast Cancer Patients, APMIS (2016)