Mutated PIGGYBAC Transposase for Stable High-Titer Cell Expression
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Solution Overview
Problem
The biopharmaceutical industry faces challenges in achieving high recombinant protein production and long-term stability of expression in host cells, particularly in CHO cells, which are crucial for producing biologics, due to high R&D costs and long development lead times.
Innovation Solution
Engineering PIGGYBAC® transposases with specific amino acid substitutions at positions 147, 176, 221, 247, 429, 533, and 573 to enhance stability and titer of recombinant proteins, using engineered nucleic acid sequences to improve transfection efficiency and protein expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If wild type PIGGYBAC transposase is used for transfecting cells, then transfection can be achieved, but the stability of transposase expression and recombinant protein titer remain insufficient
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the PIGGYBAC transposase at specific positions (147, 176, 221, 247, 429, 533, and 573) to optimize its stability and functionality. These sequence modifications result in enhanced transposase stability and improved recombinant protein titers compared to wild type, directly resolving the contradiction between reliability and productivity.
2Duration of action of stationary object
If conventional transfection methods are used, then cells can be transfected, but long-term stability of expression is not achieved
Solution Approach 1:
The engineered PIGGYBAC transposase with modified amino acid sequences achieves both long-term expression stability and high productivity. The sequence optimizations at specific positions enable sustained transposase activity and stable integration of transgenes, resulting in improved long-term expression stability without compromising recombinant protein production levels.
3Loss of time
If R&D time is reduced for biopharmaceutical development, then speed-to-market improves, but expression stability may be compromised
Solution Approach 1:
The patent employs preliminary action by pre-optimizing the PIGGYBAC transposase sequence with identified beneficial mutations before actual transfection applications. This pre-engineering of the transposase ensures that subsequent transfections achieve both rapid results and stable long-term expression, effectively reducing development lead time while maintaining or improving expression stability.
Data Source
AI summary
PIGGYBAC® transposases engineered to increase stability in a cell. The engineered PIGGYBAC® transposases are useful for stably transforming cells, cell line development, genome modification, and improving titer of recombinant proteins, among other uses.