Cell Engineering Platform for Alcohol-Mediated Non-Adherent Cell Delivery
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Solution Overview
Problem
Existing methods for delivering particles and molecules into cells, particularly non-adherent cells, face challenges due to the cell membrane's selective barrier, leading to high costs, complexity, variable results, and cell damage, especially at scale.
Innovation Solution
A vector-free delivery method using an alcohol-containing aqueous solution to permeabilize the cell membrane, combined with a closed system platform for scalable, automated, and sterile transfection of non-adherent cells, enabling efficient delivery of mRNA and RNP to primary T cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional transfection methods are used for non-adherent cells, then delivery efficiency is poor, but the process becomes more complex and costly
Solution Approach 1:
The invention changes the chemical parameters of the delivery solution by incorporating alcohol at specific concentrations (greater than 2% v/v, preferably 5-30%) to permeabilize cell membranes. This parameter change enables efficient transfection of non-adherent cells without requiring complex viral vectors or specialized equipment, thus improving reliability while reducing process complexity
Solution Approach 2:
The invention introduces an alcohol-containing aqueous solution as an intermediary substance that facilitates payload delivery across the cell membrane. This intermediary agent (alcohol) temporarily permeabilizes the membrane to allow mRNA or RNP entry, then reverses to restore membrane integrity, achieving reliable transfection through a simple chemical mediator rather than complex mechanical or biological systems
2Productivity
If conventional transfection methods are used, then cell damage occurs, but scalability is limited
Solution Approach 1:
The invention uses a rapid, short-duration alcohol exposure protocol that quickly permeabilizes cell membranes for payload delivery and then immediately restores membrane integrity. This rushed-through approach minimizes the time cells are in a vulnerable state, reducing cell damage while enabling scalable processing of large cell numbers (10^7 to 10^9 cells) in a single use
Solution Approach 2:
The invention employs a simple, disposable alcohol-containing delivery solution that can be applied and discarded without complex recovery or sterilization processes. This approach enables scalable processing by allowing high-throughput treatment of large cell populations without the need for expensive, reusable equipment or complex contamination control protocols
3Speed
If vector-free delivery is implemented, then delivery speed increases, but contamination risk increases
Solution Approach 1:
The alcohol-containing aqueous solution serves as a sterile intermediary vehicle that delivers the payload rapidly while maintaining a closed system. The solution can be prepared and stored under sterile conditions, applied quickly to cells, and then removed or neutralized, enabling fast delivery without compromising sterility
Solution Approach 2:
The invention creates a controlled, sterile environment by using an alcohol-containing aqueous solution that can be prepared and handled in a closed system. The alcohol component provides some antimicrobial properties, and the aqueous buffer maintains physiological conditions, together creating an inert-like environment that protects against contamination during rapid delivery processes
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 method achieves rapid and reliable delivery to a large number of cells, supports scalable processes, reduces contamination risks, and enhances cell survivability and throughput, while being easy to use and repeatable.
Implementation Method 1
contacting the population of cells with a volume of an aqueous solution, the aqueous solution including the payload and an alcohol at greater than 2 percent (v/v) concentration
Data Source
AI summary
The current subject matter provides a cell engineering platform of a diagnostic and clinical use scale for vector-free and/or viral delivery of payload/cargo compounds and compositions into non-adherent cells. The platform achieves delivery to a large number of cells quickly in a closed system. Related apparatus, systems, techniques, articles and compositions are also described.