Optimizing Eukaryotic Protein Secretion via Genetic Element Screening
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Solution Overview
Problem
The high cost and inefficiency of producing recombinant proteins in eukaryotic cells, particularly for complex molecules like monoclonal antibodies, due to the limitations in choosing optimal mRNA targeting elements such as signal sequences and untranslated regions, which affect protein synthesis and secretion levels.
Innovation Solution
A kit containing multiple pre-defined secretion cassettes with different combinations of genetic elements, including promoters, 5' and 3' untranslated regions, and signal sequences, is used to screen for the optimal cassette that maximizes protein synthesis and secretion in eukaryotic cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If eukaryotic cells are used to produce complex recombinant proteins like monoclonal antibodies, then the ability to perform correct post-translational modifications and protein folding is improved, but the production cost increases and efficiency decreases
Solution Approach 1:
The patent optimizes mRNA targeting elements (signal sequences, 5' and 3' UTRs) to change the parameters of protein synthesis and secretion efficiency. By selecting specific combinations of these elements, the patent achieves high-level protein production in eukaryotic cells while maintaining their post-translational modification capabilities.
Solution Approach 2:
The patent divides the secretion cassette into distinct functional segments: signal sequence (SS), 5' untranslated region (5'UTR), coding sequence (cds), 3' untranslated region (3'UTR), and poly(A) site. By independently optimizing each segment and testing various combinations, the patent identifies the most effective configuration for high-yield protein production.
2Quantity of substance
If the demand for therapeutic proteins increases, then the need for high-yield production is improved, but the complexity of the production process increases
Solution Approach 1:
The patent performs preliminary optimization of secretion cassettes by testing multiple combinations of mRNA targeting elements before large-scale production. This preliminary action identifies the optimal cassette configuration, simplifying subsequent production processes and enabling high-yield protein manufacturing.
Solution Approach 2:
The patent systematically varies parameters of mRNA targeting elements (different signal sequences from various sources, different UTR configurations) to identify combinations that maximize protein yield. This parameter optimization approach simplifies the overall production process by finding the most efficient configuration.
3Productivity
If different signal sequences are tested to optimize protein secretion, then the protein production level is improved, but the time and resources required for screening increase
Solution Approach 1:
The patent segments the screening process into systematic comparisons of specific mRNA targeting element combinations. By organizing the screening around defined cassette configurations with specific SS and UTR combinations, the patent efficiently identifies optimal sequences without exhaustive testing of all possible variations.
Solution Approach 2:
The patent tests a selected set of representative signal sequences and UTR combinations rather than all possible variations. This partial action approach identifies sufficiently optimal configurations for high-level protein production without the time and resource expenditure of complete exhaustive screening.
Data Source
Figure 1A~1B
Figure 2
AI summary
The invention relates to a kit to be used for the identification of a secretion cassette, which in eukaryotic cells gives rise to a higher level of synthesis/secretion of a protein of interest, the coding sequence of which being flanked by the first and the second set of genetic elements, respectively, compared to the other secretion cassettes present in the kit containing the coding sequence at the corresponding position, as well as to a method wherein the kit is used and the use of said kit, whereby the best secretion cassette will be obtained.