Production and evaluation of scfvs against her2 antigen for breast cancer immunotherapy

Elham Nadimi,1,*

1. Recombinant Antibody Laboratory, Department of Immunology, Shiraz University of Medical Sciences, Shiraz, IR Iran

Abstract

Introduction

Breast cancer is common cause of women death in the world. there is erbb2/her2 in 25%-30% breast cancer cases. trastuzumab and pertuzumab antibodies, are currently used for treat her2-positive breast cancer, although their clinical use is limited due to their immunogenicity. human recombinant single-chain antibodies(scfvs), which are produced by antibody engineering technologies are new and effective in cancer immunotherapy.to product specific single-chain variable fragments(scfvs) against 2‏ immunodominant her2 epitopes, to evaluate their reactivity and specificity against her2 epitopes.

Methods

E.coli bacteria, containing a phagemid with a scfv insert segment, were used to product specific high-affinity scfvs against 2 her2 epitopes, by using panning process. polymerase chain reaction(pcr) and dna fingerprinting were performed obtained from clones to product positive ones and isolate the common patterns. the production clones were evaluated via phage elisa in terms of reactivity and epitopes specificity

Results

Scfvi and scfvii, which are her2-specific with 40% and 45% frequencies, were producted against epitopes i and ii, respectively. phage-elisa finding demonstrated a significant difference in scfvs optical density(od) in reaction with related peptides and non-peptide wells. ods of 0.65 and 0.71 were obtained for scfvi and scfvii reactions with corresponding peptides, whereas ods of non-peptide wells were 0.1 and 0.13, respectively.

Conclusion

Targeted cancer therapy, which acts on a specific molecule in cancer cells, minimizes the side effects of immunotherapy. in this study, 2‏specific scfvs were selected against 2 her2 epitopes. potentially these small, high-affinity, human antibodies to be considered for breast cancer targeted therapy. however, in vitro and in vivo tests should be performed to evaluate the scfvs's antitumor effects.

Keywords

Breast cancer, her2, scfv antibodies, targeted therapy