Electroporation Apparatus Asymmetric Flow Path
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Solution Overview
Problem
Current electroporation systems for large-scale manufacturing are inefficient and costly, with low throughput and poor viability for transfecting mammalian cells, particularly NK cells, due to non-uniform electric fields and high costs of commercial products.
Innovation Solution
An electroporation apparatus with an offset input and output design, using exponentially discharging or square waveforms, and a fluid channel region between electrodes, applying multiple electrical pulses at varying field strengths and durations to achieve high efficiency and viability for cell transfection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional parallel plate electrodes are used for electroporation, then the device structure is simple, but the electric field is non-uniform causing poor transfection efficiency
Solution Approach 1:
The patent introduces an offset between the input and output positions of the cell suspension, creating an asymmetric flow path through the electroporation chamber. This asymmetric configuration ensures that cells traverse a more uniform electric field region, improving transfection efficiency without requiring complex electrode structures
Solution Approach 2:
The patent adds a spatial dimension to the cell flow path by offsetting the input and output positions. This dimensional change allows cells to move through the electric field in a trajectory that avoids regions of high field non-uniformity, achieving better transfection efficiency
2Productivity
If commercial electroporation products are used for large scale production, then the transfection capability is available, but the cost is high and throughput is low
Solution Approach 1:
The patent optimizes electroporation parameters including applying multiple electrical pulses with varying field strengths (0.3-3 kV/cm) and durations (10 μs to 10,000 μs). These parameter optimizations enable high transfection efficiency (70-90%) and cell viability (>80%) while using cost-effective, scalable equipment configurations
Solution Approach 2:
The patent employs multiple periodic electrical pulses instead of a single pulse, with controllable intervals between pulses. This periodic action increases transfection efficiency by allowing cumulative pore formation while giving cells time to recover, enabling high-throughput operation with maintained cell viability
3Manufacturing precision
If high electric field strength is applied for electroporation, then transfection efficiency improves, but cell viability decreases
Solution Approach 1:
The patent applies multiple electrical pulses with optimized intervals between them. The periodic application allows cumulative electroporation effect while providing recovery time, achieving high transfection efficiency (70-90%) while maintaining cell viability (>80%)
Solution Approach 2:
The patent uses dynamically adjustable electrical pulse parameters including variable field strength (0.3-3 kV/cm) and pulse duration (10 μs to 10,000 μs). This dynamic parameter adjustment allows optimization of the balance between transfection efficiency and cell viability based on specific cell types and applications
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 system provides improved transfection efficiency and cell viability for mammalian cells, including NK cells, by maintaining a uniform electric field and optimizing electroporation parameters, thus addressing the limitations of existing methods.
Implementation Method 1
Electroporation involves applying a controlled direct current (DC) electrical pulse to a cell for a relatively short duration of time. The electrical pulse is thought to induce a transmembrane potential that causes a reversible breakdown of the ordered structure of a cell membrane, leading to the formation of pores in the membrane.
Data Source
AI summary
Systems and methods are provided for transfecting cells, such as mammalian cells and nonmammalian cells, using an electroporation apparatus having an electroporation chamber including a first electrode, a second electrode and a path defined in the electroporation chamber. The electroporation apparatus includes a first input allowing passage of cells and cargo into the electroporation chamber and a first output allowing passage of electroporated cells from the electroporation chamber.