The Production of HPV Type 16 Pseudovirion for the Development of Pseudovirion-Based Neutralization Assay (PBNA)
Production of HPV-16 Pseudovirion for PBNA
Keywords:
HPV-16 pseudovirion production, Plasmid transfection, Pseudovirion-based neutralization assay (PBNA)Abstract
Human papillomavirus (HPV) is implicated in the majority of cervical cancer cases, which vaccination being a crucial method for preventing infection. The pseudovirion-based neutralization assay (PBNA) has been employed to determine the neutralizing antibody levels in HPV-vaccine immunized recipients. This article introduced the product of HPV-pseudovirion serotype 16 (HPV-16 pseudovirion), a technique transferred from the protocol established by the National Institutes for Food and Drug Control (NIFDC), China. Results following the transfection of p16sheLL and pcDNA3.1 eGFP plasmids into 293FT cell lines showed the production of pseudovirion containing the eGFP protein within 48 hours, with infectivity observed on the 293FT cell line. Fluoresced cells were counted using ELIPOT. The calculated pseudovirion titer, based on 300 eGPF-positive cells per 0.1 milliliter, was 4.37, with a dilution factor of 11,771 for PBNA. Additionally, the pseudovirion titer examined through TCID50 per 0.1 milliliter was 6.67, with a dilution factor of 11,585 for PBNA. Overall, the adopted method successfully produced HPV-16 pseudovirion, serving as a prototype for producing other HPV-pseudovirion serotypes and facilitating their transfer to relevant agencies to support the HPV vaccination campaign
References
Zhang X, Li S, Modis Y, Li Z, Zhang J, Xia N, et al. Functional assessment and structural basis of antibody binding to human papillomavirus capsid. Rev Med Virol 2016; 26(2): 115-28.
Burd EM. Human papillomavirus and cervical cancer. Clin Microbiol Rev 2003; 16(1): 1-17.
Egawa N, Doorbar J. The low-risk papillomaviruses. Virus Res 2017; 231: 119-27.
Brisson M, Kim JJ, Canfell K, Drolet M, Gingras G, Burger EA, et al. Impact of HPV vaccination and cervical screening on cervical cancer elimination: a comparative modelling analysis in 78 low-income and lower-middleincome countries. Lancet 2020; 395(10224): 575-90.
Wang JW, Roden RB. Virus-like particles for the prevention of human papillomavirus-associated malignancies. Expert Rev Vaccines 2013; 12(2): 129-41.
Yang DY, Bracken K. Update on the new 9-valent vaccine for human papillomavirus prevention. Can Fam Physician 2016; 62(5): 399-402.
Ma B, Roden RB, Hung CF, Wu TC. HPV pseudovirions as DNA delivery vehicles. Ther Deliv 2011; 2(4): 427-30.
Biryukov J, Meyers C. Papillomavirus infectious pathways: a comparison of systems. Viruses 2015; 7(8): 4303-25.
Tsang SH, Basu P, Bender N, Herrero R, Kemp TJ, Kreimer AR, et al. Evaluation of serological assays to monitor antibody responses to single-dose HPV vaccines. Vaccine 2020; 38(38): 5997-6006.
Nie J, Liu Y, Huang W, Wang Y. Development of a triple-color pseudovirion-based assay to detect neutralizing antibodies against human papillomavirus. Viruses 2016; 8(4): 107. (12 pages).
Sehr P, Rubio I, Seitz H, Putzker K, Ribeiro-Müller L, Pawlita M, et al. High-throughput pseudovirion-based neutralization assay for analysis of natural and vaccine-induced antibodies against human papillomaviruses. PLoS One 2013; 8(10): e75677. (11 pages).
Nie J, Huang W, Wu X, Wang Y. Optimization and validation of a high throughput method for detecting neutralizing antibodies against human papillomavirus (HPV) based on pseudovirons. J Med Virol 2014; 86(9): 1542-55.
Buck CB, Pastrana DV, Lowy DR, Schiller JT. Generation of HPV pseudovirions using transfection and their use in neutralization assays. Methods Mol Med 2005; 119: 445-62.
Buck CB, Thompson CD, Pang YY, Lowy DR, Schiller JT. Maturation of papillomavirus capsids. J Virol 2005; 79(5): 2839-46.
Buck CB, Pastrana DV, Lowy DR, Schiller JT. Efficient intracellular assembly of papillomaviral vectors. J Virol 2004; 78(2): 751-7.
Prachasuphap A, Jongpitisub K, Saengtong N, Treeyoung P, Chaiya P, Dhepakson P. Cloning and expression of human monoclonal antibody with potent binding to 2019 novel coronavirus. Bull Dept Med Sci 2020; 62(3): 155-66.
Downloads
Published
Versions
- 02-07-2024 (2)
- 30-06-2024 (1)
How to Cite
Issue
Section
License
Copyright (c) 2024 BULLETIN OF THE DEPARTMENT OF MEDICAL SCIENCES
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
