Related Experiment Video
Updated: Jun 13, 2026
13:51
The MultiBac Protein Complex Production Platform at the EMBL
Published on: July 11, 2013
15.9K
Continuous Production of Influenza VLPs Using IC-BEVS and Multi-Stage Bioreactors.
Ricardo Correia1,2, Taja Zotler3, Francisco Ferraz1,2
1Instituto de Biologia Experimental e Tecnológica (iBET), Oeiras, Portugal.
Biotechnology and Bioengineering
|January 18, 2025
Summary
Continuous production of influenza hemagglutinin-displaying virus-like particles (HA-VLPs) was achieved using the insect cell-baculovirus expression vector system (IC-BEVS). This intensified process maintained consistent HA-VLPs titers for 20 days, overcoming previous limitations.
Area of Science:
- Biotechnology and Bioprocessing
- Molecular Virology
- Cell Culture Engineering
Background:
- The insect cell-baculovirus expression vector system (IC-BEVS) is a well-established platform for biologic production.
- Process intensification, including continuous manufacturing, is a key trend in biotechnology.
- Previous continuous production efforts with IC-BEVS were limited by cell lysis and virus passage effects.
Purpose of the Study:
- To develop and implement an intensified, continuous production process for influenza hemagglutinin-displaying virus-like particles (HA-VLPs) using IC-BEVS.
- To evaluate different insect cell lines and recombinant baculovirus constructs for optimal continuous HA-VLP production.
- To assess the impact of bioreactor residence times on HA-VLP yield and quality.
Main Methods:
- A multi-stage bioreactor system was designed, comprising a cell growth bioreactor feeding three parallel production bioreactors.
- Production bioreactors were operated at distinct residence times (18, 36, and 54 hours) and continuously harvested.
- Two insect cell lines (High Five, Sf9) and two recombinant baculovirus constructs (rBACbacmid, rBACflashbac) were tested.
Main Results:
- The combination of rBACflashbac with Sf9 cells yielded consistent HA-VLPs titers (34 ± 14 HA titer/mL) and baculovirus titers (108-109 pfu/mL) over 20 days of continuous operation.
- Higher residence times correlated with increased HA-VLP expression, with 54 hours maximizing titers.
- Transmission electron microscopy confirmed the presence of HA-VLPs throughout the continuous production period.
Conclusions:
- A continuous production process for HA-VLPs using IC-BEVS was successfully implemented, demonstrating process intensification.
- The developed system overcomes limitations associated with the lytic nature of IC-BEVS and virus passage effects.
- This work establishes a foundation for integrated, continuous biomanufacturing platforms utilizing the IC-BEVS.
Keywords:
baculovirus vector designcontinuous productioninfluenza HA‐VLPsinsect cellsmulti‐stage bioreactorsMore Related Videos
10:11Modeling The Lifecycle Of Ebola Virus Under Biosafety Level 2 Conditions With Virus-like Particles Containing Tetracistronic Minigenomes
Published on: September 27, 2014
36.1K
08:10Production of High-Titer Infectious Influenza Pseudotyped Particles with Envelope Glycoproteins from Highly Pathogenic H5N1 and Avian H7N9 Viruses
Published on: January 15, 2020
8.2K
Related Concept Videos
01:14Scale-Up Processes
The scale-up of microbial fermentation processes is essential in industrial biotechnology, allowing the transition from laboratory-scale experiments to commercial-scale production while aiming to maintain product yield and quality. This process requires meticulous adjustment of equipment design, process parameters, and contamination control strategies to accommodate increasing culture volumes.At the laboratory scale, cultures are typically maintained in 1 to 10-liter glass or autoclavable...
01:27Upstream Processing
Upstream processing represents a critical phase in biomanufacturing, wherein biological systems such as microorganisms, mammalian cells, or insect cells are cultivated to produce therapeutic proteins, vaccines, enzymes, or other biologically derived products. This phase encompasses all steps from the selection and genetic manipulation of the production organism to the cultivation of cells in bioreactors under tightly controlled environmental conditions.Host Selection and Genetic OptimizationThe...
01:30Production of Pharmaceuticals
Industrial insulin production uses genetically engineered E. coli expressing a proinsulin gene controlled by a tryptophan promoter and containing a methionine linker for later cleavage. The cells also carry ampicillin resistance for selective growth. Seed cultures are stored at −80 °C and production begins by thawing a small amount to inoculate starter cultures, which are progressively scaled to a 50,000-L bioreactor. In the bioreactor, E. coli grow in nutrient-rich media under sterile, tightly...
01:23Vaccine Production
Vaccine production involves a sequence of upstream and downstream processes to generate a safe and effective immunological product. It begins with cultivating microorganisms, such as viruses or bacteria, to obtain antigenic material. For viral vaccines, mammalian host cells are grown in bioreactors and subsequently infected with the target virus. The virus replicates within the host cells, which are lysed to release viral particles. This lysate is then clarified through filtration or...