High-Throughput Production of Influenza Virus-Like Particle (VLP) Array by Using VLP-factory, a MultiBac Baculoviral Genome Customized for Enveloped VLP Expression

Methods Mol Biol. 2019:2025:213-226. doi: 10.1007/978-1-4939-9624-7_10.

Abstract

Baculovirus-based expression of proteins in insect cell cultures has emerged as a powerful technology to produce complex protein biologics for many applications ranging from multiprotein complex structural biology to manufacturing of therapeutic proteins including virus-like particles (VLPs). VLPs are protein assemblies that mimic live viruses but typically do not contain any genetic material, and therefore are safe and attractive alternatives to life attenuated or inactivated viruses for vaccination purposes. MultiBac is an advanced baculovirus expression vector system (BEVS) which consists of an engineered viral genome that can be customized for tailored applications. Here we describe the creation of a MultiBac-based VLP-factory, based on the M1 capsid protein from influenza, and its application to produce in a parallelized fashion an array of influenza-derived VLPs containing functional mutations in influenza hemagglutinin (HA) thought to modulate the immune response elicited by the VLP.

Keywords: Baculovirus expression vector system (BEVS); Cre recombinase; Cre-LoxP fusion; Hemagglutinin (HA); Influenza; MultiBac; Small-scale production; Virus-like particle (VLP).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Baculoviridae / genetics*
  • Genome, Viral / genetics*
  • Hemagglutinins / genetics
  • Hemagglutinins / metabolism
  • Orthomyxoviridae / genetics
  • Orthomyxoviridae / metabolism

Substances

  • Hemagglutinins