Plasmid Extraction via Venturi Mixing and Continuous Flow
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Solution Overview
Problem
Current methods for plasmid extraction face challenges in efficiently handling a range of quantities, with batch methods limiting scalability and continuous methods incurring significant fixed costs for smaller quantities, while also risking shearing of genomic DNA and plasmids due to mechanical stirring.
Innovation Solution
A method involving a receptacle with mechanical stirring, where an alkaline lysis solution is added to a cell suspension, followed by a neutralizing solution and a precipitating solution via multiple orifices, allowing for controlled mixing and extraction with gentle stirring, enabling high yield and purity of plasmid extraction across varying quantities without shearing issues.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If mechanical stirring is used to mix solutions in batch mode, then homogeneity of mixture is improved, but shear forces cause shearing of genomic DNA and plasmids
Solution Approach 1:
The patent replaces mechanical stirring with a Venturi effect-based mixing system. The Venturi mixer uses fluid dynamics (pressure differential created by constricted flow) to generate mixing action without mechanical moving parts, thereby eliminating shear forces that would damage DNA and plasmids while still achieving homogeneous mixing of the viscous solutions.
Solution Approach 2:
The invention employs hydraulic principles through the Venturi effect, where the flow of liquid through a constricted section creates a pressure differential that draws in and mixes additional solutions. This hydraulic mixing mechanism achieves homogeneity without the harmful mechanical shear forces of traditional stirrers.
2Ease of operation
If batch mode extraction is used, then mixing control is improved, but scalability to larger quantities is limited by container size and mixing efficiency
Solution Approach 1:
The patent implements continuous extraction by replacing the batch-wise sequential addition of solutions with continuous flow streams. Cell suspension, lysis solution, neutralizing solution, and precipitating solution all flow continuously through the Venturi mixing system, enabling scalable processing of large quantities while maintaining precise mixing control through flow rate regulation.
Solution Approach 2:
The system transitions from static batch mixing to dynamic continuous flow mixing. Flow rates of different solutions can be independently adjusted to optimize mixing at different stages, and the system can adapt to varying production volumes by simply changing flow rates rather than container sizes.
3Productivity
If continuous extraction method is used, then productivity for large quantities is improved, but fixed costs become significant for smaller quantities
Solution Approach 1:
The Venturi-based continuous extraction system is designed to be universally applicable across a wide range of production volumes. The same continuous flow system can process from small to large quantities by simply adjusting flow rates, eliminating the need for separate batch and continuous systems and making the technology cost-effective for both small and large-scale operations.
4Manufacturing precision
If batch mode extraction is used with low volumes, then purification efficiency is improved, but production on larger scale is significantly complicated
Solution Approach 1:
The continuous extraction system segments the extraction process into distinct functional zones within the flow path: lysis zone, neutralization zone, and precipitation zone. Each zone has optimized flow rates and mixing characteristics, allowing high purification efficiency to be maintained while scaling to large volumes without proportionally increasing system complexity.
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
This method achieves high extraction yield and purity of plasmids, maintaining flexibility for both small and large quantities, reducing contamination, and allowing for efficient production on a larger scale with controlled shear forces.
Implementation Method 1
The neutralising and precipitating solutions are mixed homogeneously by adapting the internal diameter of the piping, so as to locally create a Venturi effect.
Implementation Method 2
obtaining a receptacle 1 coupled to an extraction pump 8, said receptacle 1 being provided with mechanical stirring means 9
Implementation Method 3
clarifying, preferably centrifuging, said extracted mixture so as to recover the clarified supernatant, comprising said plasmid of interest
Data Source
AI summary
A method is for discontinuous plasmid extraction from a microorganism and for purifying a plasmid of interest. The method includes adding a neutralizing solution and then a precipitation solution via a plurality of orifices, under mild mechanical stirring.