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Developing and characterization of single chain variable fragment (scFv) antibody against frizzled 7 (Fzd7) receptor

Hamid Nickho1,2,3, Vahid Younesi4,5, Leili Aghebati-Maleki1,2,3

  • 1a Immunology Research Center, Tabriz University of Medical Sciences , Tabriz , Iran.

Bioengineered
|November 17, 2016
PubMed

Insights

Researchers developed targeted therapies by isolating antibody fragments against Frizzled 7 (Fzd7) to inhibit cancer cell growth. These Fzd7-targeting single-chain variable fragments (scFvs) show potential for immunotherapy in triple-negative breast cancer.

Area of Science:

  • Molecular Biology
  • Immunology
  • Oncology

Background:

  • Wnt/β-catenin signaling, mediated by Frizzled receptors, is crucial in development and cancer.
  • Overexpression of Wnt pathway genes like Fzd7 is common in malignancies.
  • Targeting Fzd7 and its ligands can inhibit cancer cell proliferation and metastasis.

Purpose of the Study:

  • To isolate single-chain variable fragments (scFvs) targeting the Frizzled 7 (Fzd7) receptor.
  • To evaluate the efficacy of these scFvs in inhibiting cancer cell growth and inducing apoptosis.
  • To explore the potential of Fzd7-targeting scFvs for breast cancer immunotherapy.

Main Methods:

  • Phage display technology using semi-synthetic human naive antibody libraries (Tomlinson I + J).
  • Panning procedures to select specific scFvs against Fzd7 extracellular domain peptides.
  • Enzyme-linked immunosorbent assay (ELISA), MTT, and annexin V assays to assess reactivity and effects on cell growth and apoptosis.

Main Results:

  • Seven scFvs specifically reactive to Fzd7 were successfully isolated after four rounds of panning.
  • The selected scFvs demonstrated significant inhibition of cell growth in MDA-MB-231 triple-negative breast cancer cells.
  • These scFvs induced cell death through apoptosis.

Conclusions:

  • Targeting Fzd7 with isolated scFvs offers a promising strategy for cancer therapy.
  • The identified Fzd7-blocking scFvs hold significant potential for the immunotherapy of breast cancer.
  • This approach could be valuable given Fzd7's critical role in tumor progression via the Wnt pathway.
Keywords:
Fzd7Wntbreast cancerphage displayscFv

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