Scalable production of HPV16 L1 protein and VLPs from tobacco leaves

Maryam Zahin, Joongho Joh, Sujita Khanal, Adam Husk, Hugh Mason, Heribert Warzecha, Shin Je Ghim, Donald M. Miller, Nobuyuki Matoba, Alfred Bennett Jenson

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Cervical cancer is the most common malignancy among women particularly in developing countries, with human papillomavirus (HPV) 16 causing 50% of invasive cervical cancers. A plant-based HPV vaccine is an alternative to the currently available virus-like particle (VLP) vaccines, and would be much less expensive.We optimized methods to express HPV16 L1 protein and purify VLPs from tobacco (Nicotiana benthamiana) leaves transfected with the magnICON deconstructed viral vector expression system. L1 proteins were extracted from agro-infiltrated leaves using a series of pH and salt mediated buffers. Expression levels of L1 proteins and VLPs were verified by immunoblot and ELISA, which confirmed the presence of sequential and conformational epitopes, respectively. Among three constructs tested (16L1d22, TPL1d22, and TPL1F), TPL1F, containing a full-length L1 and chloroplast transit peptide, was best. Extraction of HPV16 L1 from leaf tissue was most efficient (> 2.5% of total soluble protein) with a low-salt phosphate buffer. VLPs were purified using both cesium chloride (CsCl) density gradient and size exclusion chromatography. Electron microscopy studies confirmed the presence of assembled forms of HPV16 L1 VLPs. Collectively; our results indicated that chloroplast-targeted transient expression in tobacco plants is promising for the production of a cheap, efficacious HPV16 L1 VLP vaccine. Studies are underway to develop plant VLPs for the production of a cervical cancer vaccine.

Original languageEnglish (US)
Article numbere0160995
JournalPLoS One
Volume11
Issue number8
DOIs
StatePublished - Aug 1 2016

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Tobacco
uterine cervical neoplasms
tobacco
Virus-Like Particle Vaccines
virus-like particle vaccines
Uterine Cervical Neoplasms
leaves
Buffers
Proteins
buffers
proteins
chloroplasts
Salts
Human papillomavirus 16
vaccines
salts
Papillomavirus Vaccines
Cancer Vaccines
cesium
Size exclusion chromatography

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Zahin, M., Joh, J., Khanal, S., Husk, A., Mason, H., Warzecha, H., ... Jenson, A. B. (2016). Scalable production of HPV16 L1 protein and VLPs from tobacco leaves. PLoS One, 11(8), [e0160995]. https://doi.org/10.1371/journal.pone.0160995

Scalable production of HPV16 L1 protein and VLPs from tobacco leaves. / Zahin, Maryam; Joh, Joongho; Khanal, Sujita; Husk, Adam; Mason, Hugh; Warzecha, Heribert; Ghim, Shin Je; Miller, Donald M.; Matoba, Nobuyuki; Jenson, Alfred Bennett.

In: PLoS One, Vol. 11, No. 8, e0160995, 01.08.2016.

Research output: Contribution to journalArticle

Zahin, M, Joh, J, Khanal, S, Husk, A, Mason, H, Warzecha, H, Ghim, SJ, Miller, DM, Matoba, N & Jenson, AB 2016, 'Scalable production of HPV16 L1 protein and VLPs from tobacco leaves', PLoS One, vol. 11, no. 8, e0160995. https://doi.org/10.1371/journal.pone.0160995
Zahin, Maryam ; Joh, Joongho ; Khanal, Sujita ; Husk, Adam ; Mason, Hugh ; Warzecha, Heribert ; Ghim, Shin Je ; Miller, Donald M. ; Matoba, Nobuyuki ; Jenson, Alfred Bennett. / Scalable production of HPV16 L1 protein and VLPs from tobacco leaves. In: PLoS One. 2016 ; Vol. 11, No. 8.
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