Plug-and-Display: decoration of Virus-Like Particles via isopeptide bonds for modular immunization

Scientific Reports
Karl D BruneMark Howarth

Abstract

Virus-like particles (VLPs) are non-infectious self-assembling nanoparticles, useful in medicine and nanotechnology. Their repetitive molecularly-defined architecture is attractive for engineering multivalency, notably for vaccination. However, decorating VLPs with target-antigens by genetic fusion or chemical modification is time-consuming and often leads to capsid misassembly or antigen misfolding, hindering generation of protective immunity. Here we establish a platform for irreversibly decorating VLPs simply by mixing with protein antigen. SpyCatcher is a genetically-encoded protein designed to spontaneously form a covalent bond to its peptide-partner SpyTag. We expressed in E. coli VLPs from the bacteriophage AP205 genetically fused to SpyCatcher. We demonstrated quantitative covalent coupling to SpyCatcher-VLPs after mixing with SpyTag-linked to malaria antigens, including CIDR and Pfs25. In addition, we showed coupling to the VLPs for peptides relevant to cancer from epidermal growth factor receptor and telomerase. Injecting SpyCatcher-VLPs decorated with a malarial antigen efficiently induced antibody responses after only a single immunization. This simple, efficient and modular decoration of nanoparticles should accele...Continue Reading

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Datasets Mentioned

BETA
KU302810
KU302811

Methods Mentioned

BETA
protein folding
transmission electron microscopy
electrophoresis
enzyme-linked immunosorbent assay
ELISA
fluorescence microscopy
PCR
size-exclusion chromatography
Protein Assay
gel filtration

Software Mentioned

ImageJ
SpyCatcher
QuantityOne

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