Journal Article

DKFZ-2026-00723

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png In Silico Design and Subsequent Expression of Human Papillomavirus-16 and -18 L1 Vaccine Antigens in Broccoli.

Batool, N. ; Ahsan, K. ; Qadeer, K. ; Fajar, A. ; Farid, A. ; Sameeullah, M. ; Ijaz, F. ; Malik, M. S. ; Tariq, F. A. ; Lössl, A. G. ; Müller, M.DKFZ* ; Waheed, M. T.

2026
MDPI Basel

Abstract: Background: Cervical carcinoma remains a widespread cancer worldwide, primarily caused by persistent infection with high-risk human papillomavirus (HPV). HPV types 16 and 18 account for approximately 70% of cervical cancer cases. Although prophylactic HPV vaccines are commercially available, their high cost and reliance on expensive expression platforms limit their accessibility in developing countries. Objectives: This study aimed to develop a cost-effective, plant-based HPV vaccine candidate by expressing capsomeric HPV-16 and HPV-18 L1 antigens in Brassica oleracea (broccoli). Methods: Modified L1 from HPV types 16 and 18 were designed to retain capsomeric assembly and fused with heat-labile enterotoxin B subunit (LTB). Immunoinformatics analyses were used to assess antigenicity, epitope distribution, and structural characteristics. Codon-optimized genes were cloned using Gateway® technology and expressed in broccoli via Agrobacterium-mediated transformation. Transgenic plants were validated by PCR and qRT-PCR. Protein accumulation was quantified, and immunogenicity was evaluated in mice. Results: PCR and qRT-PCR confirmed the stable integration of two copies of the LTB-L1 transgenes in broccoli plants. Western blotting detected L1 protein at ~56.5 kDa, indicating the cleavage of the LTB-L1 fusion protein. The correct folding of L1 capsomeres was verified by antigen-capture ELISA. The recombinant proteins accumulated to approximately 0.33% and 0.35% of total soluble protein for HPV-16 and HPV-18, respectively. The immunization of mice with transgenic L1 induced significant humoral immune responses, comparable to those elicited by purified VLPs. Conclusions: The results demonstrate broccoli as a promising platform for the expression of immunogenic HPV L1 capsomeres and highlight its potential for the development of affordable, plant-based HPV vaccines.

Keyword(s): Agrobacterium-mediated transformation ; Gateway® cloning ; HPV ; LTB-L1 ; cervical cancer ; immunoinformatics

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Contributing Institute(s):

1. Tumorvirus-spez. Vakzinierungsstrategie (D335)

Research Program(s):

1. 314 - Immunologie und Krebs (POF4-314) (POF4-314)

Appears in the scientific report 2026

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Database coverage:
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