Design and Purification of Tag/Catcher AP205-Based Capsid Virus-Like Particle Vaccines

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearchpeer-review

Capsid virus-like particles (cVLPs), assembled from viral coat proteins, are used as therapeutic cargo delivery vehicles as well as molecular scaffolds for display of vaccine antigens. A versatile vaccine platform has been developed based on the Acinetobacter phage AP205 cVLP, which has been shown to significantly improve antigen-specific antibody responses. This modular cVLP platform exploits a split-protein (Tag/Catcher) conjugation system to enable high-density, unidirectional antigen display. Accordingly, protein antigens can be independently expressed and quality-checked prior to conjugation to pre-assembled cVLPs. Here, we describe considerations for the design of vaccine antigens with genetically fused split-protein (Tag or Catcher) binding partners and provide protocols for the expression and purification of corresponding Tag- or Catcher-AP205 cVLPs from E.coli. Finally, we describe a generic protocol for the formulation and quality assessment of experimental/pre-clinical AP205 cVLP-based vaccines.

Original languageEnglish
Title of host publicationTherapeutic Proteins : Methods and Protocols
PublisherHumana Press
Publication date2024
Pages127-141
ISBN (Print)978-1-0716-3468-4
ISBN (Electronic)978-1-0716-3469-1
DOIs
Publication statusPublished - 2024
SeriesMethods in Molecular Biology
Volume2720
ISSN1064-3745

Bibliographical note

Publisher Copyright:
© The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024.

    Research areas

  • AP205, Catcher, cVLP, Density ultracentrifugation, Tag, Virus-like particles

ID: 378966950