Abstract
Introduction: The scFvs (single-chain fragment variable) are popular recombinant antibody formats that are produced by advanced molecular biology and genetic engineering techniques. Moreover, HER2-ECD recognized as a suitable target for a wide range of cancer cells. The current study was accomplished for isolation of specific scFv antibodies. Methods: To isolate specific scFv fragments from phage library, phage display technology was carried out by coating HER2 extra cellular domain (HER2-ECD) protein onto an immuno tube. The resultant scFv antibodies were expressed in Escherichia coli TG1. The recombinant products were purified by metal affinity chromatography against His-tag. These scFvs were characterized for binding to HER2-ECD by western blot, biosensor surface resonance and flow cytometry assays. Results: After three rounds of panning against HER2-ECD protein yielded three different scFv fragments that confirmed by subsequent DNA sequencing. The kinetic results indicated that all of isolated scFvs had binding tendency to HER2-ECD on micro molar to nano molar range. Western blot and flow cytometry results demonstrated that all of isolated scFvs have had binding tendency to HER2-ECD. Conclusion: Phage display is a powerful tool for isolate promising scFv clones that bind specifically to the HER2 receptor, a cell surface of breast cancer antigen.
Original language | English |
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Pages (from-to) | 19-37 |
Number of pages | 19 |
Journal | Iranian Journal of Obstetrics, Gynecology and Infertility |
Volume | 17 |
Issue number | 92 |
State | Published - 2014 |
Keywords
- Antibody engineering
- Biosensor
- Flow cytometry
- HER2 receptor
- Phage display