Related Experiment Video
Updated: Jan 21, 2026

Methanol Independent Expression by Pichia Pastoris Employing De-repression Technologies
Published on: January 23, 2019
Assessing virus like particles formation and r-HBsAg aggregation during large scale production of recombinant
Seyed Nezamedin Hosseini1, Taravat Sarvari2, Ghazal Bashiri2
1Department of recombinant Hepatitis B Vaccine, Production and Research Complex, Pasteur Institute of Iran, Iran.
Abstract:
The aggregation of recombinant proteins in the different stages of purification leads to the loss of a considerable portion of target protein and reduction in the process efficiency. As the active HBsAg used in Hepatitis B vaccine production is in the form of virus-like particle (VLP), therefore the time and stages at which the VLP assembling happened through the process would be important. The aim of this study was to explore the product aggregation during different stages of large scale production of rHBsAg in Pichia pastoris at production unit of the Pasteur Institute of Iran. Dynamic light scattering (DLS) and transmission electron microscopy (TEM), and also size exclusion-high-performance liquid chromatography (SE-HPLC) were carried out on samples taken from each downstream processes steps to determine the rate of VLPs formation as the desired product and the aggregated form at each stage of the purification. Based on the results, it was found that VLPs formation started at the acid precipitation stage and reached up to 80% at the thermal treatment stage. The ultrafiltration, ion exchange chromatography and immunoaffinity chromatography stages were disclosed to have the highest contribution in the formation of VLP (virus like particle) 22 nm.
Related Concept Videos
Precipitate Formation and Particle Size Control
The obtained precipitate should be either a pure substance of known composition or easily converted to one by a simple process, such as ignition or drying. In addition, the precipitate should be insoluble and easily filterable. In general, filterability...
Viral Recombination
Recombinant DNA
What are Viruses?
Factors Affecting Dissolution: Particle Size and Effective Surface Area
Effect of Hepatic Disease on Pharmacokinetics: Pathophysiologic Assessment and Liver Function Test

