Newsletter | September 24, 2018

09.24.18 -- Scalable Processes For Adenovirus Vector Production

 
Scalable Process For Adenovirus Production
 

This article describes a process for adenovirus production, from upstream cell culture to downstream purification, using modern tools and technologies. To demonstrate robustness of the upstream process, virus production in suspension-adapted cells was performed in both rocking and stirred-tank bioreactors. The downstream process was carefully optimized to meet stringent regulatory demands on product purity and quality.

Adenovirus Production In A Single-Use Bioreactor System
 

Adenovirus-based vectors have been widely evaluated as vaccine delivery systems in preclinical and clinical studies of various infectious diseases. Adenovirus has also been extensively explored as a viral vector for gene therapy and as an oncolytic virus. This article describes the production of adenovirus in HEK293 cells cultured in HyClone CDM4HEK293 culture medium.

Determination Of Adenovirus Concentration
 

The performance of two assays for determination of adenovirus concentration based on surface plasmon resonance is demonstrated here. In one assay, virus particles were allowed to bind via the fiber protein to recombinant human coxsackie adenovirus receptor protein immobilized on a Biacore sensor chip. In a second assay, Factor X protein was immobilized on a Biacore sensor chip to allow binding of virus particles via the virus hexon protein.

Downstream Process Development For Efficient Purification Of Adenovirus
 

Here we describe the development of two effective downstream processes for purification of adenovirus from cell culture harvest. After screening of 11 different anion exchange chromatography adsorbents in small scale, a resin and a membrane adsorber were selected for the capture step. Process conditions were optimized, and polishing steps were included for further reduction of impurities to meet regulatory requirements.

Optimization Of Midstream Cell Lysis And Virus Filtration Steps In An Adenovirus Purification Process
 

In this article, the screening of detergents for release of adenovirus from HEK293 host cells to replace the traditionally used detergent Triton is described. Consumables and conditions that provided the highest virus purity and recovery were selected as a suggestion for a midstream process for adenovirus recovery from a 10 L cell culture, to provide process material ready for the subsequent downstream purification step.