Mammalian Cell Line Producing Soluble CSFV-E2 Antigen
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Solution Overview
Problem
Current methods for producing E2 recombinant antigen for classical swine fever virus are costly, and existing vaccines struggle to distinguish between immune responses from vaccination and natural infection, necessitating a low-cost production method for effective vaccination and diagnostic purposes.
Innovation Solution
A mammalian cell line, specifically the Chinese hamster ovary cell line CCC_E2_5E12, is used to stably express and produce soluble E2 recombinant antigen, which is then utilized to create a porcine subunit vaccine composition, significantly increasing production yield and reducing costs through mammalian cell expression systems.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If insect cells are used to produce E2 recombinant antigen, then the antigen can be produced, but the production cost remains high
Solution Approach 1:
The patent replaces expensive insect cell systems with a cheaper transient transfection system using eukaryotic cells (HEK293 or CHO cells). The transient expression system allows for cost-effective production by using disposable transfection reagents and cells that can be cultured in standard mammalian cell culture conditions, eliminating the need for expensive insect cell infrastructure while achieving high antigen yield
Solution Approach 2:
The patent optimizes multiple parameters including transfection method (Lipofectamine 2000), cell density, medium composition, and incubation conditions to maximize antigen production in the transient expression system. By adjusting these parameters, the system achieves high yields of soluble E2 antigen at lower cost compared to conventional insect cell production
2Reliability
If modified live vaccine is used, then vaccination is cheap and effective, but it is impossible to distinguish immune response from vaccination or natural infection
Solution Approach 1:
The patent produces purified E2 envelope protein as a subunit vaccine, separating the antigen into its specific functional component. This segmented approach allows the antigen to retain immunogenicity while eliminating viral genetic material, enabling safe subunit vaccination that can be distinguished from live virus infection through diagnostic testing
Solution Approach 2:
The patent extracts and purifies only the E2 envelope protein from the virus, removing all other viral components including genetic material. This extracted subunit antigen maintains the ability to induce protective immunity while allowing clear distinction between vaccine-induced and infection-induced immune responses through specific antibody detection
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method achieves high production yields of soluble E2 recombinant antigen, reducing production costs and enabling effective vaccination with a single dose, while improving the safety and efficacy of CSFV-E2 protein subunit vaccines by producing a high proportion of soluble protein.
Implementation Method 1
a mammalian cell line stably expressing soluble E2 recombinant antigen of classical swine fever virus in mass
Data Source
AI summary
The present disclosure relates to a mammalian cell line and a method for producing soluble E2 recombinant antigen of classical swine fever virus using the same. A mammalian cell expression system with different cell passage numbers can stably express a large amount of soluble CSFV-E2 recombinant protein, effectively reducing the production cost of CSFV-E2 recombinant protein, and then applied to the production of CSFV-E2 protein subunit vaccines.