B-cell Co-cultivation with Optimized EL4-B5 Feeder Density
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Solution Overview
Problem
Current methods for obtaining monoclonal antibodies, such as hybridoma technology and co-cultivation with EL4-B5 feeder cells, face limitations in efficiency and specificity, particularly in achieving high frequencies of antigen-binding and IgG-positive wells, especially when EL4-B5 cell densities are not optimized.
Innovation Solution
Co-cultivating B-cells with EL4-B5 feeder cells obtained from cultivations at densities exceeding 1,000,000 cells/ml up to 1,500,000 cells/ml, using a feeder mix that includes cytokines like IL-1β, TNF-α, IL-2, IL-10, and Staphylococcus aureus strain Cowan's cells, to enhance antigen binding and antibody production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If EL4-B5 cells are cultivated at lower densities (conventional method), then the co-cultivation process is simpler to control, but the frequency of antigen-binding and IgG-positive wells is reduced
Solution Approach 1:
The patent applies parameter changes by optimizing the EL4-B5 cell density parameter to a specific range (1,000,000 to 1,500,000 cells/ml) during cultivation. This quantitative parameter optimization directly increases the frequency of antigen-binding and IgG-positive wells while maintaining controllable process complexity, resolving the contradiction between productivity and process simplicity.
2Productivity
If EL4-B5 cell density is increased to improve antibody production, then antibody productivity increases, but control over the co-cultivation system becomes more difficult
Solution Approach 1:
The patent defines a precise parameter range (1,000,000 to 1,500,000 cells/ml) for EL4-B5 cell density that maximizes antibody productivity while maintaining system reliability. This optimized parameter range ensures high antibody production without compromising the controllability and stability of the co-cultivation system.
Solution Approach 2:
The patent implements feedback control by monitoring and adjusting EL4-B5 cell density within the optimal range during cultivation. This feedback mechanism ensures that the system maintains reliable control while achieving high antibody productivity, preventing both under-cultivation (low productivity) and over-cultivation (loss of control).
Data Source
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AI summary
Herein is reported a method for co-cultivating one or more B-cells comprising the step of incubating the one or more B-cells with EL4-B5 cells, whereby the EL4-B5 cells have been obtained/are from a cultivation of EL4-B5 cells that has a cell density of from 600,000 cells/ml up to 1,500,000 cells/ml.