Translational fusion of chloroplast-expressed human papillomavirus type 16 L1 capsid protein enhances antigen accumulation in transplastomic tobacco

Paolo Lenzi, Nunzia Scotti, Fiammetta Alagna, Maria L. Tornesello, Andrea Pompa, Alessandro Vitale, Angelo De Stradis, Luigi Monti, Stefania Grillo, Franco M. Buonaguro, Pal Maliga, Teodoro Cardi

Research output: Contribution to journalArticle

Abstract

Human Papillomavirus (HPV) is the causal agent of cervical cancer, one of the most common causes of death for women. The major capsid L1 protein self-assembles in Virus Like Particles (VLPs), which are highly immunogenic and suitable for vaccine production. In this study, a plastid transformation approach was assessed in order to produce a plant-based HPV-16 L1 vaccine. Transplastomic plants were obtained after transformation with vectors carrying a chimeric gene encoding the L1 protein either as the native viral (L1 v gene) or a synthetic sequence optimized for expression in plant plastids (L1pt gene) under control of plastid expression signals. The L1 mRNA was detected in plastids and the L1 antigen accumulated up to 1.5% total leaf proteins only when vectors included the 5prime;-UTR and a short N-terminal coding segment (Downstream Box) of a plastid gene. The half-life of the engineered L1 protein, determined by pulse-chase experiments, is at least 8 h. Formation of immunogenic VLPs in chloroplasts was confirmed by capture ELISA assay using antibodies recognizing conformational epitopes and by electron microscopy.

Original languageEnglish
Pages (from-to)1091-1102
Number of pages12
JournalTransgenic Research
Volume17
Issue number6
DOIs
Publication statusPublished - Dec 2008

Keywords

  • HPV16
  • L1
  • Nicotiana tabacum
  • Plant vaccines
  • Plastid transformation
  • Tobacco

ASJC Scopus subject areas

  • Biotechnology
  • Genetics
  • Agronomy and Crop Science
  • Animal Science and Zoology

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