Bahareh Moazen1,2, Amir Zarrinhaghighi1,2, Foroogh Nejatollahi1,2. 1. Shiraz HIV/AIDS Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. 2. Recombinant antibody laboratory, Department of Immunology, Shiraz University of Medical Sciences, Shiraz, Iran.
Abstract
BACKGROUND: CD90, a membrane-associated glycoprotein is a marker used to identify mesenchymal stem cells (MSCs). Recent studies have introduced CD90, which induces tumorigenic activity, as a cancer stem cell (CSC) marker in various malignancies. Blocking CD90 activity with anti-CD90 monoclonal antibodies enhanced anti-tumor effects. To date, highly specific antibody single-chain variable fragments (scFvs) have been isolated against various targets and showed promising results in cancer immunotherapy. METHODS: A phage antibody was produced from a scFv library using M13KO7 helper phage. The phage library was panned against a CD90 epitope. To select specific clones, PCR and DNA fingerprinting were performed and common patterns were identified. The panning results were confirmed by phage ELISA. RESULTS: Of 20 clones selected after panning, 16 shared identical fingerprints. One clone from this group reacted specifically with the epitope in phage ELISA. The average absorbance of wells coated with the CD90 peptide was significantly greater than that of wells containing no peptide (p=0.03). CONCLUSION: Currently, recombinant antibodies are used not only as highly specific detection tools, but due to their specific characteristics, are applied in targeted cancer therapies. The anti-CD90 scFv selected in this study has the potential to be used to detect MSCs and target CSCs and offers promising strategies for treatment of various cancers.
BACKGROUND: CD90, a membrane-associated glycoprotein is a marker used to identify mesenchymal stem cells (MSCs). Recent studies have introduced CD90, which induces tumorigenic activity, as a cancer stem cell (CSC) marker in various malignancies. Blocking CD90 activity with anti-CD90 monoclonal antibodies enhanced anti-tumor effects. To date, highly specific antibody single-chain variable fragments (scFvs) have been isolated against various targets and showed promising results in cancer immunotherapy. METHODS: A phage antibody was produced from a scFv library using M13KO7 helper phage. The phage library was panned against a CD90 epitope. To select specific clones, PCR and DNA fingerprinting were performed and common patterns were identified. The panning results were confirmed by phage ELISA. RESULTS: Of 20 clones selected after panning, 16 shared identical fingerprints. One clone from this group reacted specifically with the epitope in phage ELISA. The average absorbance of wells coated with the CD90 peptide was significantly greater than that of wells containing no peptide (p=0.03). CONCLUSION: Currently, recombinant antibodies are used not only as highly specific detection tools, but due to their specific characteristics, are applied in targeted cancer therapies. The anti-CD90 scFv selected in this study has the potential to be used to detect MSCs and target CSCs and offers promising strategies for treatment of various cancers.
Entities:
Keywords:
CD90; Cancer stem cell; Mesenchymal stem cell; Single chain antibody
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