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Inhibitors of Virion Maturation and Assembly

Inhibitors of Virion Maturation and Assembly

As part of their replication cycle, certain viruses synthesize long precursor proteins called polyproteins within infected host cells. In human immunodeficiency virus (HIV), two major polyproteins are produced: Gag and Gag-Pol. The Gag polyprotein supplies the structural components of the virus, while Gag-Pol includes essential viral enzymes such as reverse transcriptase, integrase, and protease. After synthesis, these polyproteins move to the host cell membrane, where they assemble into an...

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Related Experiment Video

Updated: Jun 23, 2026

Expression and Purification of Virus-like Particles for Vaccination

Expression and Purification of Virus-like Particles for Vaccination

Published on: June 2, 2016

Downstream process design for Gag HIV-1 based virus-like particles.

Elianet Lorenzo¹, Laia Miranda^1,2, Francesc Gòdia¹

¹Departament d'Enginyeria Química Biològica i Ambiental, Universitat Autònoma de Barcelona, Bellaterra, Spain.

Biotechnology and Bioengineering

|May 6, 2023

Summary

This study developed an effective purification process for virus-like particles (VLPs) used in vaccines. The optimized method achieves high purity and concentration, meeting regulatory standards for VLP production.

Keywords:

HIV-1 Gag-eGFP VLP chromatography clarification downstream processing nanoparticle quantification

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Area of Science:

Biotechnology
Vaccine Manufacturing
Downstream Processing

Background:

Virus-like particles (VLPs) are crucial for modern vaccine development.
Purifying VLPs is challenging due to similar characteristics with host cell extracellular vesicles.
Efficient VLP purification is essential for vaccine safety and efficacy.

Purpose of the Study:

To compare various downstream processing technologies for VLP purification.
To develop an optimized purification train for HIV-1 Gag-eGFP VLPs.
To evaluate the yield, purity, and contaminant removal efficiency of different purification steps.

Main Methods:

Investigated clarification (depth filtration), intermediate (tangential flow filtration/multimodal chromatography), capture (ion exchange, heparin affinity, hydrophobic interaction chromatography), and polishing (size exclusion chromatography) steps.
Assessed recovery, purity, and contaminant elimination at each stage.
Implemented a complete purification train based on optimal step performance.

Main Results:

Achieved a final concentration of 1.40 × 10^10 VLPs/mL with 64% purity after polishing.
Host cell DNA and protein levels met regulatory standards.
Overall recovery of the purification process was 38%.

Conclusions:

Developed a scalable purification process for HIV-1 Gag-eGFP VLPs.
The optimized process effectively removes contaminants while maintaining VLP integrity.
This purification strategy is suitable for industrial VLP vaccine production.