On-Column T Cell Selection and Stimulation Without Detachment Reagents
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Solution Overview
Problem
Existing cell therapy methods require separate selection and stimulation steps for generating suitable cell populations, which prolong the manufacturing process and can contaminate cells, causing stress and requiring additional wash steps.
Innovation Solution
A method for on-column stimulation of T cells using an oligomeric stimulatory reagent containing anti-CD3 and anti-CD28 antibodies, which immobilizes T cells on a stationary phase and delivers a stimulatory signal without additional agents, allowing collection by gravity flow within 24 hours.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If separate selection and stimulation steps are used, then cell selection and stimulation can be achieved, but the manufacturing process is prolonged
Solution Approach 1:
The patent combines cell selection and stimulation into a single integrated process using a stationary phase that simultaneously performs both functions. The stationary phase contains selection agents for isolating target cells and stimulation agents for activating them, eliminating the need for separate steps and reducing manufacturing time while maintaining effectiveness.
Solution Approach 2:
The stationary phase is designed as a multi-functional component that performs multiple operations: cell selection through specific binding, cell stimulation through immobilized activators, and potential cell release mechanisms. This universal component replaces multiple separate reagents and steps, streamlining the manufacturing process.
2Reliability
If selection agents and competition reagents are used, then cell selection is achieved, but cells are contaminated with selection-related particles
Solution Approach 1:
The patent extracts or eliminates the need for competition reagents and detachment agents from the process. By using a stationary phase where cells are selected and stimulated in place, the method removes harmful substances that would otherwise contaminate the cell product, requiring only minimal wash steps for purification.
3Object-generated harmful factors
If additional wash steps and media exchange are performed, then cell purification is achieved, but cell stress increases and processing time extends
Solution Approach 1:
The stationary phase enables cells to be selected and stimulated in their culture medium without requiring transfer to new media or extensive washing. The system serves itself by maintaining cells in a supportive environment throughout the process, minimizing stress from media changes and handling while still achieving purification through the selective binding properties of the stationary phase.
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 reduces processing time and minimizes cell handling, generating a composition of stimulated T cells efficiently without additional contamination or stress.
Implementation Method 1
the stationary phase includes a selection agent that specifically binds to a selection marker on the surface of one or more T cells or subset thereof
Implementation Method 2
the oligomeric stimulatory reagent includes one or more stimulatory agent including (i) a first stimulatory agent that is an anti-CD3 antibody, and (ii) a second stimulatory agent that is an anti-CD28 antibody
Implementation Method 3
within 24 hours of initiating incubation, collecting one or more of the plurality of T cells from the stationary phase by gravity flow
Data Source
AI summary
Provided herein are methods for selecting and stimulating a plurality of cells in a sample of cells using column chromatography, and collecting the cells without using additional steps or reagents to facilitate detachment of the cells from the column. In some aspects, the methods provided herein reduce the time needed to generate a population of selected and stimulated cells useful for genetic engineering, and ultimately, cell therapy, compared to existing methods. Also provided are articles of manufacture and apparatus thereof.