Production of Well-Characterized Virus-like Particles in an Escherichia coli-Based Expression Platform for Preclinical Vaccine Assessments

Wahome, N.; Cooper, A.; Thapa, P.; Choudhari, S.; Gao, F.P.; Volkin, D.B.; Middaugh, C.Russell.

Methods in Molecular Biology 1404: 437-457


ISSN/ISBN: 1064-3745
PMID: 27076315
DOI: 10.1007/978-1-4939-3389-1_29
Accession: 058631856

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In this chapter we demonstrate a method to produce virus-like particles (VLPs) from Escherichia coli. Standard bacterial protocols are used for the cloning, transformation, and expression of the protein subunits. A two-step protein purification method is highlighted: one step based on separating soluble proteins with ion-exchange affinity chromatography and a second polishing step using size-exclusion columns to isolate VLP species. The ensuing VLPs can be characterized with a variety of biophysical techniques including ultraviolet (UV)-visible spectroscopy for protein quantification, dynamic light scattering for size distribution determination, and transmission electron microscopy to ascertain size and morphology.