Adenovirus Production Fed-Batch Culture HEK293 BalanCD
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Solution Overview
Problem
Current methods for producing adenovirus vectors face challenges in yield and scalability, particularly for non-AdHu5 serotypes and diverse vaccine antigens, with existing processes being complex, costly, and difficult to transfer to new facilities, and achieving high yields is limited by cell density effects and interference from viral-replication-inhibitory factors.
Innovation Solution
A fed batch culture method using HEK293 cells with BalanCD medium and BalanCD HEK293 Feed, where cells are infected at high densities and fed with 5% of the medium volume every 24-48 hours, allowing for improved adenovirus production without the need for perfusion or medium exchange, utilizing HEK293-T-REx cells and an antigen-repressing promoter system to enhance yields.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If perfusion-based processes are used to achieve high yields, then productivity is improved, but device complexity and ease of manufacture deteriorate
Solution Approach 1:
The invention extracts and eliminates the perfusion step from the manufacturing process. By using a simplified batch process without perfusion, the patent achieves high virus yields (up to 2×10^14 VP/L) while removing the complex equipment and operational procedures associated with perfusion systems, making the process easier to manufacture and transfer to new facilities
Solution Approach 2:
The invention segments the manufacturing process into distinct, manageable stages: cell growth phase, infection phase, and harvest phase. Each phase has optimized parameters that can be independently controlled, eliminating the need for continuous perfusion while maintaining high productivity through staged process optimization
2Productivity
If cell density is increased to improve productivity, then virus yield is improved, but harmful factors increase
Solution Approach 1:
The invention applies preliminary action by using a defective viral genome that cannot replicate independently. This defective genome is designed to hijack the cellular machinery for producing full-length viral genomes without being able to replicate on its own, thereby preventing the accumulation of harmful viral-replication-inhibitory factors while maintaining high productivity
Solution Approach 2:
The invention introduces an intermediary mechanism where a defective viral genome acts as a template that requires cellular enzymes and factors to produce functional viral genomes. This intermediary approach allows high-level virus production while the defective nature of the template prevents it from generating harmful replication intermediates
3Adaptability or versatility
If diverse vaccine antigens are used to improve adaptability, then versatility is improved, but manufacturing precision deteriorates
Solution Approach 1:
The invention extracts the problematic replicating viral genome component and replaces it with a defective genome that cannot undergo genetic recombination. This extraction eliminates the source of genetic instability while preserving the ability to produce diverse vaccine antigens through the defective genome's ability to direct production of full-length viral genomes carrying various transgenes
Data Source
AI summary
The invention relates to a method for preparing an adenovirus comprising a) providing a host cell in a medium capable of supporting growth of said host cell b) contacting said host cell with an adenovirus c) incubating to allow infection of said cell by said adenovirus d) incubating to allow production of adenovirus by said host cell wherein said host cell is, or is derived from, a HEK293 cell, and wherein the medium comprises BalanCD HEK293. The invention also relates to adenovirus produced, and to compositions comprising said adenovirus.