Scfv of Nimotuzumab can enter EGFR-overexpressing cancer cells
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Seyedeh Roghayeh Hamidi,1 Yaghoub Safdari,2,* Mehdi Sheikh-Arabi,3
1. Department of Medical Biotechnology, Faculty of Advanced Technologies in Medicine, Golestan University of Medical Sciences,
2. Golestan Research Center of Gastroenterology and Hepatology, Golestan University of Medical Sciences
3. Medical cellular and molecular research center, Golestan University of Medical Sciences
- Introduction: Introduction
The epidermal growth factor receptor (EGFR) is a glycoprotein tyrosine kinase receptor which is the main factor in controlling of the cell growth and viability. Since this receptor is overexpressed in many carcinomas, it is regarded as an ideal target for cancer therapy and imaging. Anti-EGFR monoclonal antibodies can be engaged to specifically deliver small molecules with therapeutic or diagnostic properties to EGFR-overexpressing cancer cells. Therefore, in the current study, we evaluated the ability of single-chain Nimotuzumab molecules in recognizing and entering EGFR-overexpressing cancer cells.
- Methods: Methods
Amino acid sequence of Nimotuzumab variable domains (VH and VL) was obtained from the protein data bank (PDB). Nine arginine residues (called 9-R) was added to N-terminus of VH domain to form a scFv in VH-linker-VL format. After gene synthesis and cloning into pET22b (+), the recombinant was expressed and conjugated to FITC. The conjugate was tested on A-431 (EGFR-overexpressed cells) and MCF-7 (EGFR low-expressed cells) cancer cells. Scfv molecules lacking 9R segment was used as a control.
- Results: Results
Both the molecules were found to enter the cancer cells (A431 Cells) in an efficient manner and emit signals. No obvious difference was found between CPP-containing and CPP-lacking molecules in entering cancer cells.
- Conclusion: Conclusion
9R-nimotuzumab ScFv is able to recognize and enter the EGFR-overexpressing cancer cells. CPP is not an essential component for cellular internalization.
- Keywords: EGFR, 9R-scFv, scFv, A-431, MCF-7.
