Volume 1 - Supplement Issue 1: Abstracts of International Tehran Breast Cancer Congress
Multidiscip Cancer Investig 2017, 1 - Supplement Issue 1: Abstracts of International Tehran Breast Cancer Congress: 0-0 |
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Development of anti-FZD7 scFv Antibody and Evaluation of its Apoptotic Effect on Breast Cancer Cells
Abstract: (4845 Views)
Introduction: Frizzled family receptor 7 (FZD7) is one of the 10 members of Frizzled
receptors. It interacts with Wnt ligands to activate canonical wnt signaling, which turns on
different downstream transcription factors essential for modulating cellular proliferation,
polarity, and differentiation. Altered expression of FZD7 receptor is associated with development
and progression of many cancers including breast cancer. An effective targeted
therapy for breast cancer through modulating ligand-receptor interaction may involve the
use of antibodies to antagonize FZD7. ScFvs (single-chain fragment variable) have provided
an alternative to full-length monoclonal antibodies (mAbs) in diagnostic and therapeutic
applications.
Materials and Methods: A phage antibody library of scFv was used and selection of specific
scFvs were performed by 4 rounds of panning process against an immunodominant epitope
of FZD7, followed by PCR and fingerprinting of the selected clones. ELISA was used
to confirm the specificity of the clones. Apoptotic effects of the selected scFv on MDAMB-
231 cell line were assessed by annexin V/PI assay after 24 h and 48 h.
Results: A specific scFv with the frequency of 35% was isolated which produced positive
ELISA with the corresponding epitope. After 24h treatment with the selected scFv,
MDA-MB-231 cells showed 48.7% apoptotic cell death (Annexin V+/PI−). However, this
amount increased to 81.6% following 48h treatment with scFv.
Conclusions: Due to unique apoptotic properties of selected scFv including human origin,
high affinity and specificity, this agent has been applied in cancer immunotherapy. The
specific anti-FZD7 scFv selected in this study has the potential to be used for inhibiting
wnt signaling pathway in breast cancer cells.
receptors. It interacts with Wnt ligands to activate canonical wnt signaling, which turns on
different downstream transcription factors essential for modulating cellular proliferation,
polarity, and differentiation. Altered expression of FZD7 receptor is associated with development
and progression of many cancers including breast cancer. An effective targeted
therapy for breast cancer through modulating ligand-receptor interaction may involve the
use of antibodies to antagonize FZD7. ScFvs (single-chain fragment variable) have provided
an alternative to full-length monoclonal antibodies (mAbs) in diagnostic and therapeutic
applications.
Materials and Methods: A phage antibody library of scFv was used and selection of specific
scFvs were performed by 4 rounds of panning process against an immunodominant epitope
of FZD7, followed by PCR and fingerprinting of the selected clones. ELISA was used
to confirm the specificity of the clones. Apoptotic effects of the selected scFv on MDAMB-
231 cell line were assessed by annexin V/PI assay after 24 h and 48 h.
Results: A specific scFv with the frequency of 35% was isolated which produced positive
ELISA with the corresponding epitope. After 24h treatment with the selected scFv,
MDA-MB-231 cells showed 48.7% apoptotic cell death (Annexin V+/PI−). However, this
amount increased to 81.6% following 48h treatment with scFv.
Conclusions: Due to unique apoptotic properties of selected scFv including human origin,
high affinity and specificity, this agent has been applied in cancer immunotherapy. The
specific anti-FZD7 scFv selected in this study has the potential to be used for inhibiting
wnt signaling pathway in breast cancer cells.
Select article type: Original/Research Article |
Subject:
Materials Science and Nanomedicine
Received: 2017/10/28 | Accepted: 2017/10/28 | ePublished: 2017/10/28
Received: 2017/10/28 | Accepted: 2017/10/28 | ePublished: 2017/10/28
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