ArticleApplied biochemistry and biotechnology2024
Production of a Ribosome-Displayed Mouse scFv Antibody Against CD133, Analysis of Its Molecular Docking, and Molecular Dynamic Simulations of Their Interactions.
Article in Applied biochemistry and biotechnology, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed, 7 citations in OpenAlex.
- Bioinformatic analysis and characterization of the anti-CD20 scFv-conjugated FeIranian journal of basic medical sciences · 2026Article
- In Vitro Selection of Antibodies by Ribosome Display.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Advances in Techniques for the Structure and Functional Optimization of Therapeutic Monoclonal Antibodies.Biomedicines · 2025Review
- Application of Nanomaterial-Based Sonodynamic Therapy in Tumor Therapy.Pharmaceutics · 2024Review
- Design, Expression, and Binding Interactions Study of the Recombinant Engineered IL-6R.Iranian journal of pharmaceutical research : IJPRArticle
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
The pentaspan transmembrane glycoprotein CD133, prominin-1, is expressed in cancer stem cells in many tumors and is promising as a novel target for the delivery of cytotoxic drugs to cancer-initiating cells. In this study, we prepared a mouse library of single-chain variable fragment (scFv) antibodies using mRNAs isolated from mice immunized with the third extracellular domain of a recombinant CD133 (D-EC3). First, the scFvs were directly exposed to D-EC3 to select a new specific scFv with high affinity against CD133 using the ribosome display method. Then, the selected scFv was characterized by the indirect enzyme-linked immunosorbent assay (ELISA), immunocytochemistry (ICC), and in silico analyses included molecular docking and molecular dynamics simulations. Based on ELISA results, scFv 2 had a higher affinity for recombinant CD133, and it was considered for further analysis. Next, the immunocytochemistry and flow cytometry experiments confirmed that the obtained scFv could bind to the CD133 expressing HT-29 cells. Furthermore, the results of in silico analysis verified the ability of the scFv 2 antibody to bind and detect the D-EC3 antigen through key residues employed in antigen-antibody interactions. Our results suggest that ribosome display could be applied as a rapid and valid method for isolation of scFv with high affinity and specificity. Also, studying the mechanism of interaction between CD133's scFv and D-EC3 with two approaches of experimental and in silico analysis has potential importance for the design and development of antibody with improved properties.
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