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All Journal HAYATI Journal of Biosciences
Firdaus, Moh Egy Rahman
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Agriculture, Biological Sciences & Forestry Earth & Planetary Sciences

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Cloning, Expression, and Bioinformatics Modeling of Human Papillomavirus Type 52 L1/L2 Chimeric Protein in Escherichia coli BL21 (DE3) Ikramullah, Muh. Chaeril; Mustopa, Apon Zaenal; Wibawa, Tri; Hertati, Ai; Umami, Rifqiyah Nur; Ratna, Lita Tri; Irawan, Shasmita; Firdaus, Moh Egy Rahman; Darusman, Huda Salahudin
HAYATI Journal of Biosciences Vol. 31 No. 5 (2024): September 2024
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.31.5.891-902

Abstract

Human papillomavirus (HPV) L1 major capsid protein generates a highly immunogenic virus like particles (VLPs), which have been used as the main component of its prophylactic vaccine. However, the neutralizing antibodies against L1 VLPs are mostly type specific and may not be effective to prevent infection from different strains of HPV. On the other hand, HPV L2 minor capsid protein has low antigenic variation, thus can induce cross-neutralization. This study aims to obtain HPV 52 L1/L2 chimeric protein, which is designed based on HPV type 52 as one of the most circulated high-risk types in Indonesia, to develop a broad-spectrum HPV vaccine. Substitution of HPV 52 H4 helix L1 region with an HPV 52 L2 epitope was carried out using overlap extension PCR. HPV 52 L1/L2 chimeric gene was constructed into pET-SUMO expression vector and expressed in Escherichia coli BL21 (DE3). Bioinformatics modeling suggested that L2 epitope was located inside of the loop region in monomer form, and on the contrary, it was located outside of the pentamer surface. Furthermore, B cell and T cell epitopes predictions were conducted using Immune Epitope Database (IEDB) analysis. The B cell epitopes prediction revealed eleven potential epitopes, whereas the T cell epitopes prediction showed seven potential epitopes for each MHC class I and MHC class II. This study showed that HPV 52 L1/L2 chimeric protein has the potential to induce cross-neutralizing antibodies and can be developed as a promising candidate for a new HPV vaccine.
Truncation on N-Terminal Hydrophobic Domain of L1 Major Capsid Protein of Human Papillomavirus Type 52 Enhances Its Expression in Hansenula polymorpha Arifah, Rosyida Khusniatul; Firdaus, Moh Egy Rahman; Chairunnisa, Sheila; Irawan, Shasmita; Ekawati, Nurlaili; Irawan, Herman; Nurfatwa, Maritsa; Hertati, Ai; Swasthikawati, Sri; Novianti, Ela; Mustafawi, Wike Zahra; Nur Umami, Rifqiyah; Mustopa, Apon Zaenal
HAYATI Journal of Biosciences Vol. 32 No. 4 (2025): July 2025
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.32.4.1062-1072

Abstract

Human papillomavirus (HPV) infection is the main cause of cervical cancer. The administration of the HPV prophylactic vaccine, which is commonly produced based on HPV L1 major capsid protein, significantly reduces the incidence of cervical cancer. However, the coverage of the HPV vaccination program is often hindered due to its relatively high cost. This study aimed to evaluate the impact of N-terminal hydrophobic domain truncation on the expression of L1 major capsid protein of HPV type 52 in Hansenula polymorpha. The truncation enhanced the yield of L1 protein expression compared with the full length, which was confirmed by Western blot and ELISA. Furthermore, the truncated L1 protein formed virus-like particles (VLPs), which were confirmed by transmission electron microscopy (TEM). Bioinformatics analysis showed that the truncated L1 protein was more soluble compared with the full length, possibly increasing the protein expression. These findings could pave the way for the development of a more cost-effective HPV type 52 L1 protein production in H. polymorpha to be used as a VLP-based prophylactic vaccine.
Co-Authors Ai Hertati, Ai Apon Zaenal Mustopa Arifah, Rosyida Khusniatul Chairunnisa, Sheila Ekawati, Nurlaili Huda Shalahudin Darusman Ikramullah, Muh. Chaeril Irawan, Herman Irawan, Shasmita Mustafawi, Wike Zahra Novianti, Ela Nur Umami, Rifqiyah Nurfatwa, Maritsa Ratna, Lita Tri Rifqiyah Nur Umami, Rifqiyah Nur Sri Swasthikawati, Sri Tri Wibawa