Evaluation and analysis of synthetic HIV-1 V3 peptides incorporation with C3d adjuvant and vector design to increase the peptide expression
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Somayeh Zare,1,* Ali Salehnia sammak,2
1. High institute for research and education in transfusion medicine, Blood transfusion research center, Iran
2. Islamic azad university, department of microbiology,rasht, iran
- Introduction: One of the main sites for the production of neutralizing antibody against HIV-1 is the Glycoprotein 120 V3 region. According to the previous studies, the C4-V3 T303-E22C and the C-PRILGPG-C sequences are designed and produced as synthetic peptides that showed a suitable potential in triggering immunogenic properties against HIV. This study aims to investigate the variability of the second and third structures of these designed synthetic peptides after adding C3d adjuvant for increasing the level of immunogenicity. Additionally, a suitable vector has been designed for expressing the fusion proteins.
- Methods: After designing different arrangements of synthetic peptides with C3d adjuvant and linker, we have used Phyre2 (intensive format) and Raptorx software for homology modeling and predicting the second and third structures of fusion proteins. The stability and half-life Evaluation of designed peptide sequences applied by Protparam and Rampage software. For determining the secondary structure of RNAs obtained by expressing the nucleic acid units of our designed peptide sequences and its ∆G amount the M Fold software has been used. The location of fusion protein sequences and the type of expression host determined via PSort Prediction software.
- Results: The prediction of the second and third structures of fusion proteins showed the junction of C3d adjuvant to the beginning of the C-PRILGPG-C sequence (without linker) and the end of the C4-V3 T303-E22C (with a linker) had more similarity with the primary structure of these proteins.
- Conclusion: Due to the placement pattern of synthetic peptide sequences at the beginning and end of the C3d adjuvant peptide, the possibility of simultaneous binding of these two peptides to the adjuvant was possible without affecting the primary structures of both synthetic peptides. Additionally, our findings from Protparam analysis revealed that for expressing the C4-V3 T303-E22C fusion protein the mammalian Eukaryotic cell host is needed while expressing the C-PRILGPG-C fusion protein is suitable with both Prokaryotic and Eukaryotic cell host. Eukaryotic expression vector pGA was designed to express the fusion of proteins of this study.
- Keywords: Keywords: HIV1, Antibodies, Neutralizing, Adjuvant, Peptides
