Bioorthogonal Strategy for Bioprocessing of Specific-Site-Functionalized Enveloped Influenza-Virus-Like Particles

Sofia B. Carvalho, João M. Freire, Mafalda G. Moleirinho, Francisca Monteiro, Diana Gaspar, Miguel A R B Castanho, Manuel J T Carrondo, Paula M. Alves, Gonçalo J L Bernardes, Cristina Peixoto

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Virus-like particles (VLPs) constitute a promising platform in vaccine development and targeted drug delivery. To date, most applications use simple nonenveloped VLPs as human papillomavirus or hepatitis B vaccines, even though the envelope is known to be critical to retain the native protein folding and biological function. Here, we present tagged enveloped VLPs (TagE-VLPs) as a valuable strategy for the downstream processing and monitoring of the in vivo production of specific-site-functionalized enveloped influenza VLPs. This two-step procedure allows bioorthogonal functionalization of azide-tagged nascent influenza type A hemagglutinin proteins in the envelope of VLPs through a strain-promoted [3 + 2] alkyne-azide cycloaddition reaction. Importantly, labeling does not influence VLP production and allows for construction of functionalized VLPs without deleterious effects on their biological function. Refined discrimination and separation between VLP and baculovirus, the major impurity of the process, is achieved when this technique is combined with flow cytometry analysis, as demonstrated by atomic force microscopy. TagE-VLPs is a versatile tool broadly applicable to the production, monitoring, and purification of functionalized enveloped VLPs for vaccine design trial runs, targeted drug delivery, and molecular imaging.

Original languageEnglish
Pages (from-to)2386-2399
Number of pages14
JournalBioconjugate Chemistry
Volume27
Issue number10
DOIs
Publication statusPublished - 19 Oct 2016

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