Sphere-Packing Lattice Electroporation for Multiplexed Delivery
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Solution Overview
Problem
Traditional electroporation systems face inefficiencies due to high current requirements, environmental distortions, and limitations in delivering multiple types of exogenous materials, leading to low cell viability and single-plex delivery of molecules.
Innovation Solution
A sphere-packing lattice composition using uniform lattice-forming beads and reagent bundles that self-assemble into a crystalline structure, allowing for increased concentration of exogenous materials near cells and simultaneous delivery of multiple reagents through electrical pulses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional electroporation systems use high current input to deliver exogenous material into cells, then transformation efficiency may be improved, but excessive heat generation and medium heating occur leading to reduced cell viability
Solution Approach 1:
The invention segments the delivery of exogenous material by using multiple separate electroporation pulses instead of a single high-current pulse. The first pulse delivers the primary payload while the second pulse delivers additional material, dividing the total energy input into discrete temporal segments that prevent excessive heat accumulation in the medium.
Solution Approach 2:
The invention employs periodic action by using a sequence of electroporation pulses with specific timing intervals. The first pulse is followed by a recovery period, then a second pulse is applied. This periodic structure allows the medium to cool between pulses while still achieving cumulative transformation efficiency, directly addressing the heat generation problem.
2Productivity
If traditional electroporation systems increase the concentration of exogenous material to improve delivery, then transformation efficiency improves, but electrical conductivity of the medium increases leading to excessive Joule heating
Solution Approach 1:
The invention segments the exogenous material delivery into two separate pulses rather than delivering all material at once in high concentration. The first pulse delivers initial material at a manageable concentration, and the second pulse delivers additional material after the medium has recovered, avoiding the need to maintain high overall concentration that would increase conductivity and Joule heating.
Solution Approach 2:
The first electroporation pulse serves as a preliminary action that delivers initial exogenous material and prepares cells for subsequent uptake. This preliminary delivery allows the system to then deliver additional material in a second pulse without requiring excessively high concentrations in either pulse, thereby controlling medium conductivity and reducing Joule heating while still achieving high overall transformation efficiency.
3Adaptability or versatility
If traditional electroporation systems use single-plex delivery of payload molecules, then delivery simplicity is maintained, but the ability to deliver multiple types of exogenous material simultaneously is limited
Solution Approach 1:
The invention uses periodic action with multiple electroporation pulses to achieve multiplexed delivery. The first pulse delivers one type of exogenous material while the second pulse delivers a different type, enabling the system to deliver multiple reagents sequentially. This temporal separation through periodic pulsing allows versatile multiplexed delivery without requiring complex simultaneous multi-channel delivery systems.
Solution Approach 2:
The invention segments the multiplexed delivery function into separate temporal components by using distinct electroporation pulses for different payload types. Rather than attempting to deliver multiple materials simultaneously through a single complex pulse, the system segments delivery into sequential pulses, each optimized for specific material types, thereby achieving adaptability for multiplexed delivery while keeping individual pulse parameters relatively simple.
4Productivity
If traditional electroporation systems operate in automated multi-module cell processing environments, then throughput is improved, but environmental distortions such as electric field distortion and local pH variation increase
Solution Approach 1:
The invention segments the electroporation process into multiple controlled pulses rather than relying on a single high-intensity pulse. This segmentation reduces the peak environmental distortions (electric field distortion, local pH variation) that occur during each individual pulse, while the cumulative effect of multiple pulses maintains high transformation efficiency. This approach enables reliable operation in automated multi-module environments where maintaining stable environmental conditions across multiple processing modules is challenging.
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
Improves transformation efficiency by increasing the delivery of exogenous materials without heating the medium, enhancing cell viability and enabling multiplexed reagent delivery.
Implementation Method 1
a sphere-packing lattice composition comprising lattice-forming spherical hydrogel beads, reagent bundles and cells
Implementation Method 2
Electroporation, also known as electropermeabilization, substantially increases cell membrane permeability in the presence of a pulsed electric field
Implementation Method 3
The sphere-packing lattice utilizes lattice-forming beads that are uniform in size and that self-assemble into a crystalline-like lattice
Implementation Method 4
Because exogenous material may increase electrical conductivity (as in the case of, e.g., DNA), the upper threshold of total exogenous material that can be added to an electroporation device is determined by the amount of current/Joule heating that cells can withstand
Data Source
AI summary
The present disclosure provides a sphere-packing lattice electroporation device configured for use as a stand-alone unit or in an automated multi-module cell processing environment and configured to decrease cell processing time and cell survival. The sphere-packing lattice utilizes lattice-forming beads that are uniform in size and that self-assemble into a crystalline-like lattice.