1. Molecular dynamics simulation and purification of chimeric L1/L2 protein from human papillomavirus type 52 expressed in Escherichia coli BL21 (DE3).
- Author
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Al Adawiah R, Zaenal Mustopa A, Budiarti S, Nur Umami R, Hertati A, Irawan H, Ikramullah MC, Arwansyah A, Mamangkey J, Kartikasari I, and Salahudin Darusman H
- Subjects
- Humans, Capsid Proteins immunology, Capsid Proteins chemistry, Capsid Proteins isolation & purification, Capsid Proteins genetics, Recombinant Fusion Proteins immunology, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins isolation & purification, Recombinant Fusion Proteins biosynthesis, Oncogene Proteins, Viral immunology, Oncogene Proteins, Viral chemistry, Oncogene Proteins, Viral genetics, Oncogene Proteins, Viral isolation & purification, Human Papillomavirus Viruses, Alphapapillomavirus, Escherichia coli genetics, Escherichia coli metabolism, Molecular Dynamics Simulation
- Abstract
The available prophylactic vaccines for human papillomavirus (HPV) in the market are only effective against specific types of HPV, rendering them ineffective for other types of HPV infections. The objective of this research is to investigate the stability of the recombinant protein constructed, namely chimeric L1/L2 protein from HPV type 52, with improved cross-neutralization ability. The 3D model, predicted using Alphafold, Robetta, I-Tasser, and refined with Galaxy Refinement, is validated using Ramachandran plot analysis. The stability is verified through molecular dynamics simulations, considering parameters such as RMSD, RMSF, Rg, and SASA, where stable conditions are observed. The chimeric L1/L2 protein from HPV type 52 is purified using affinity chromatography, and the His-tag is cleaved using SUMO protease to obtain pure chimeric protein with the size of ~ 55 kDa. Western blot analysis confirms binding to anti-L1 HPV type 52 polyclonal antibody. The obtained vaccine candidate can be utilized as an effective prophylactic vaccine against HPV.
- Published
- 2024
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