Cell Engineering Platform Using Ethanol Permeabilization
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Solution Overview
Problem
Current cell engineering technologies face challenges in efficiently and consistently delivering molecules into cells due to limitations in size and charge, and existing methods like viral vectors and electroporation are costly, complex, and result in variable and inconsistent results.
Innovation Solution
A cell engineering platform that uses a solution-based delivery method with a permeabilization agent to reversibly permeabilize cell membranes, allowing for the efficient delivery of payloads across cell membranes, including the use of an aqueous solution with ethanol to enhance delivery efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional intracellular delivery methods (viral vectors, electroporation) are used, then delivery efficiency can be achieved, but the process becomes costly and complex with variable results
Solution Approach 1:
The patent changes the chemical parameters of the delivery solution by incorporating ethanol at greater than 2% (v/v) concentration, which fundamentally alters the membrane permeability characteristics and enables consistent payload delivery without complex equipment
Solution Approach 2:
The patent employs disposable pods containing the delivery solution with permeabilization agents and payloads, replacing expensive reusable equipment like electroporation devices and viral vector systems, achieving clinical-grade transfection at lower cost and complexity
2Quantity of substance
If large scale cell engineering platforms are used to obtain sufficient viable cells, then cell quantity is sufficient, but operating costs increase and greater quantities of target cells and reagent media are required
Solution Approach 1:
The patent changes the delivery mechanism by using ethanol-based permeabilization solutions that work effectively at smaller cell quantities, eliminating the need for large-scale platforms and reducing operating costs while maintaining sufficient viable cell output
3Reliability
If manual methods are used to generate experimental data from reversibly permeabilizing cells, then experimental quality can be maintained, but the process is time-consuming and manually intensive
Solution Approach 1:
The patent implements self-service through automated pod filling systems where the platform automatically fills pods with cell-mixture and first medium, and automatically discharges medium through filters, eliminating manual intervention while maintaining experimental quality and dramatically increasing productivity
4Productivity
If atomized delivery solution is applied to cells on filter, then delivery efficiency is enhanced, but precise control of solution application is required
Solution Approach 1:
The patent uses an atomizer as an intermediary device that converts the delivery solution into fine droplets for uniform distribution onto cells on the filter, enhancing delivery efficiency while the controller acts as another intermediary to precisely regulate the atomization process and solution flow
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 platform achieves reliable and consistent delivery of payloads into cells with high viability and expression, enabling clinical-grade transfection and reducing costs and complexity by automating the cell poration process, suitable for smaller scale cell processing and experimental designs.
Implementation Method 1
A delivery platform includes an atomizer; and a pod holder configured to receive the pod. One or more of the following features can be included in any feasible combination. For example, a delivery solution that contains a permeabilization agent and a payload can be sprayed onto the cells deposited on the filter.
Implementation Method 2
filling a pod of a cell engineering platform with a mixture of cells and a first medium; and discharging the first medium from the pod through a filter, leaving the cells deposited on the filter. The pod includes a filter plate and an upper portion forming a well for holding cells and media.
Implementation Method 3
A cell engineering platform that uses a solution-based delivery method with a permeabilization agent to reversibly permeabilize cell membranes, allowing for the efficient delivery of payloads across cell membranes
Implementation Method 4
The filling of the pod can be performed automatically with a pump and a controller.
Implementation Method 5
Discharging the first medium from the pod can be performed by supplying a vacuum to the bottom of the pod.
Implementation Method 6
The pod can be agitated. The resuspended cells can be extracted from the pod.
Data Source
AI summary
The current subject matter provides a cell engineering platform for vector-free and/or viral delivery of payload/cargo compounds and compositions into cells. The platform achieves delivery to cells quickly and in an easy to use manner. Related apparatus, systems, techniques, articles and compositions are also described.