Isolation and Characterization of A Panel of Anti-HER2 Scfv Molecules by Phage Display Technology

Shahrokh Ghovvati, Mojtaba Tahmoorespur, Mohammad Reza Nassiri, Ahmad Reza Bahrami, Hessam Dehghani, Hossein Borghaei, Gregory Adams

Research output: Contribution to journalArticlepeer-review

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 languageEnglish
Pages (from-to)19-37
Number of pages19
JournalIranian Journal of Obstetrics, Gynecology and Infertility
Volume17
Issue number92
StatePublished - 2014

Keywords

  • Antibody engineering
  • Biosensor
  • Flow cytometry
  • HER2 receptor
  • Phage display

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