Optimization, characterization, comparison of self-assembly VLP of capsid protein L1 in yeast and reverse vaccinology design against human papillomavirus type 52

Abstract

Background Vaccination is the one of the agendas of many countries to reduce cervical cancer caused by the Human papillomavirus. Currently, VLP-based vaccine is the most potent vaccine against HPV, which could be produced by a variety of expression systems. Our study focuses on a comparison of recombinant protein expression L1 HPV52 using two common yeasts, Pichia pastoris and Hansenulapolymorpha that have been used for vaccine production on an industrial scale. We also applied bioinformatics approach using reverse vaccinology to design alternative multi-epitope vaccines in recombinant protein and mRNA types. Results Our study found that P.pastoris relatively provided higher level of L1 protein expression and production efficiency compared to H.polymorpha in a batch system. However, both hosts showed self-assembly VLP formation and stable integration during protein induction. The vaccine we have designed exhibited high immune activation and safe in computational prediction. It is also potentially suitable for production in a variety of expression systems. Conclusion By monitoring the overall optimization parameter assessment, this study can be used as the basis reference for large-scale production of the HPV52 vaccine.

Expression and scale-up production of recombinant human papillomavirus type 52 L1 protein in methylotrophic yeast Hansenula polymorpha

“…However, the copy number is not necessarily proportional to the protein titer. 12 Moreover, recombinant protein overexpression might be damaging to host cell development and cause protein burden, which impacts protein production. 23 , 24 Meanwhile, HIP (Hansenula Integration Plasmid) which possesses an expression cassette was stably integrated into the genome of H. polymorpha and constantly expressed HPV type 52 L1 protein.…”

“…The genome was prepared with an amount of 5 ng and amplified for 40 cycles using specific primers with three-step amplification. 12 …”

“…A comparison was made between the expression of HPV type 52 L1 protein in P. pastoris and H. polymorpha using flask-scale measurements. 12 In continuity with the previous study, our study aims to optimize the production of the HPV type 52 L1 protein in the H. polymorpha expression system, a methylotrophic yeast, by utilizing the pHIPX4 Hansenula plasmid. The study encompassed many upstream development activities, including cloning and expression, investigation of clone copy quantity and stability, and medium optimization.…”

Precise Therapy Using the Selective Endogenous Encapsidation for Cellular Delivery Vector System

“…Researchers showed that self-assembled VLP formation and stable integration during protein induction were successful in both yeasts. These expression systems are used for vaccine production in the industry; however, the production effectiveness of desired proteins differs among yeasts [ 59 ].…”

Expression, purification, and characterization of self-assembly virus-like particles of capsid protein L1 HPV 52 in Pichia pastoris GS115

“…HPV infects epithelial cells involving the skin, respiratory mucosa, or genital tract. High-risk HPV is the type of HPV that causes malignant tumors to lead to cancer containing HPV 16,18,31,33,45,51,and 52 [7]. Data from the Agency for Health Research and Development, Indonesian Ministry of Health, reported that HPV has the highest prevalence in Indonesia, namely HPV 16, 18, and 52, where HPV type 52 is a highrisk HPV with the highest percentage of 18.5% [8].…”