Amphotericin B Culture Supplement for Influenza Virus Yield
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Solution Overview
Problem
Current methods for producing influenza vaccines face challenges such as contamination, variable quality, and low virus titers in primary diploid cell cultures, and unsuitability of Vero cells for large-scale production due to restricted growth and low virus titers, necessitating the development of new cultivation methods to increase yield and quality.
Innovation Solution
The use of amphotericin B as a culture supplement for the propagation of influenza A and B viruses, which enhances virus growth and infectivity, even at low multiplicity of infection, and can be used in combination with trypsin to support multicycle growth patterns.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If Vero cells are used for influenza virus production, then certification and standardization are improved, but virus growth and yield are restricted
Solution Approach 1:
The patent changes the chemical composition parameters of the culture medium by adding amphotericin B (at concentrations of 0.1-10 μg/ml) and trypsin (at concentrations of 1-100 μg/ml) to the standard Vero cell culture medium. This parameter modification enables influenza viruses to replicate efficiently in certified Vero cells, resolving the contradiction between using certified cells and achieving high virus yields.
2Ease of manufacture
If primary diploid cell cultures are used, then ease of preparation and sensitivity are improved, but contamination risk and variable quality increase
Solution Approach 1:
The patent employs disposable certified Vero cell lines that can be rapidly produced and standardized, replacing the need for complex primary diploid cell preparations. These certified cells provide consistent quality and low contamination risk while maintaining ease of preparation through standardized protocols.
Solution Approach 2:
By modifying the culture medium parameters with amphotericin B and trypsin, the patent achieves consistent high-quality virus production in certified Vero cells, eliminating the variable quality associated with primary diploid cells while maintaining preparation simplicity.
3Productivity
If repeated addition of trypsin is performed, then multicycle growth pattern is restored, but labor and time consumption increase
Solution Approach 1:
The patent incorporates trypsin directly into the culture medium at the beginning of the experiment (preliminary action), eliminating the need for repeated manual additions during the cultivation process. This preliminary incorporation maintains the multicycle growth pattern while significantly reducing labor and time requirements.
Solution Approach 2:
By including trypsin in the culture medium from the start, the patent ensures continuous presence of the enzyme throughout the virus cultivation process, maintaining consistent multicycle growth without interruption or repeated manual intervention, thus saving time and labor.
Data Source
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AI summary
The use of macrolide polyene antibiotics or derivatives or analogues thereof as culture supplement for the propagation of virus is described. Further pharmaceutical compositions comprising a virus and a macrolide polyene antibiotic or a derivative or analogue thereof and methods for using of macrolide polyene antibiotics for transfection and infection of cells as well as the use of macrolide poylene antibiotics for the isolation of virus from clinical samples are disclosed.