In silico Analysis, Molecular Docking, Molecular Dynamic, Cloning, Expression and Purification of Chimeric Protein in Colorectal Cancer Treatment
| Autor: | Mohammad Reza Zinatizadeh, Seyed Rouhollah Miri, Ghanbar Mahmoodi Chalbatani, Peyman Kheirandish Zarandi, Hassan Dana, Fereidoon Memari, Reza Rasoolzadeh, Vahid Marmari, Elahe Gharagouzloo |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Pharmacology biology In silico Pharmaceutical Science Chimeric gene Computational biology Fusion protein Epitope law.invention 03 medical and health sciences 030104 developmental biology 0302 clinical medicine Plasmid law 030220 oncology & carcinogenesis Drug Discovery MHC class I biology.protein Recombinant DNA Gene |
| Zdroj: | Drug Design, Development and Therapy. 14:309-329 |
| ISSN: | 1177-8881 |
| DOI: | 10.2147/dddt.s231958 |
| Popis: | Introduction Colorectal cancer (CRC) is a type of cancer in humans that leads to high mortality and morbidity. CD166 and CD326 are immunoglobulins that are associated with cell migration. These molecules are included in tumorigenesis of CRC and serve a great marker of CRC stem cells. In the present study, we devised a novel chimeric protein including the V1-domain of the CD166 and two epitopes of CD326 to use in diagnostic or therapeutic applications. Methods In silico techniques were launched to characterize the properties and structure of the protein. We have predicted physicochemical properties, structures, stability, MHC class I binding properties and ligand-receptor interaction of this chimeric protein by means of computational bioinformatics tools and servers. The sequence of chimeric gene was optimized for expression in prokaryotic host using online tools and cloned into pET-28a plasmid. The recombinant pET28a was transformed into the E. coli BL21DE3. Expression of recombinant protein was examined by SDS-PAGE and Western blotting. Results The designed chimeric protein retained high stability and the same immunogenicity as of the original proteins. Bioinformatics data indicated that the epitopes of the synthetic chimeric protein might induce B-cell- and T-cell-mediated immune responses. Furthermore, a gene was synthesized using the codon bias of a prokaryotic expression system. This synthetic gene expressed a bacterial expression system. The recombinant protein with molecular weights of 27kDa was expressed and confirmed by anti-his Western blot analysis. Conclusion The designed recombinant protein may be useful as a CRC diagnostic tool and for developing a protective vaccine against CRC. |
| Databáze: | OpenAIRE |
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