P-Glycoprotein Cell Selection for Cost-Effective Adoptive Therapy
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Solution Overview
Problem
Existing methods for isolating T cells for adoptive cell therapy based on mitochondrial membrane potential are slow and expensive, making them not readily amenable to clinical application.
Innovation Solution
A method for isolating p-glycoprotein (Pgp) positive cells using cytotoxic drugs, phototoxic compounds, physical or metabolic stresses, and antibody-based techniques to selectively enrich or deplete Pgp-positive cells, followed by genetic modification to express chimeric antigen receptors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If T cells are sorted based on mitochondrial membrane potential using TMRM staining and flow sorting, then functionally robust T cells with favorable metabolic features can be isolated, but the process becomes slow and expensive
Solution Approach 1:
The patent replaces expensive, time-consuming flow sorting with a simple, inexpensive magnetic bead-based sorting system that can be performed in a standard laboratory setting. The method uses commercially available magnetic beads conjugated to antibodies against cell surface markers, eliminating the need for specialized flow cytometry equipment and reducing costs while maintaining sorting effectiveness.
Solution Approach 2:
The patent extracts the essential sorting function from complex flow cytometry systems and implements it through a simplified magnetic bead-based approach. By removing unnecessary complexity and focusing on the core separation mechanism, the method achieves rapid sorting without requiring expensive equipment or specialized facilities.
2Reliability
If T cells are sorted based on mitochondrial membrane potential using TMRM staining and flow sorting, then functionally robust T cells with favorable metabolic features can be isolated, but the process becomes expensive
Solution Approach 1:
The patent replaces expensive, time-consuming flow sorting with a simple, inexpensive magnetic bead-based sorting system that can be performed in a standard laboratory setting. The method uses commercially available magnetic beads conjugated to antibodies against cell surface markers, eliminating the need for specialized flow cytometry equipment and reducing costs while maintaining sorting effectiveness.
3Ease of manufacture
If conventional cell surface marker sorting is used to identify T cells, then the process is simple and clinically feasible, but cells with long-term survival capacity may be missed
Solution Approach 1:
The patent combines conventional cell surface marker sorting with metabolic parameter assessment by using magnetic beads conjugated to antibodies against both surface markers and metabolic indicators. This dual-parameter sorting approach identifies T cells with both appropriate surface phenotype and favorable metabolic features, ensuring long-term survival capacity while maintaining clinical feasibility.
Solution Approach 2:
The patent employs composite magnetic bead particles that carry multiple antibody types, allowing simultaneous sorting based on multiple parameters including cell surface markers and metabolic features. This composite approach enables comprehensive cell selection without requiring multiple separate sorting steps.
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
Enriches cells suitable for adoptive cell therapy efficiently and cost-effectively, enhancing their survival and antitumor immunity, and provides compositions for treating cancer and infections.
Implementation Method 1
Pgp-positive cells were isolated by a number of techniques known in the art, such as magnetic activated cell sorting (MACS), in addition to conventional flow sorting. In some embodiments, the Pgp-positive cells are genetically modified.
Implementation Method 2
In some embodiments, the Pgp-positive cells are genetically modified to express at least one chimeric antigen receptor, T cell receptor, chimeric T cell receptor, synthetic immune receptor, TRuCTM or ArtemisTM T cell platform, so as to obtain genetically modified Pgp-positive T cells, NK cells, stem cells, and/or mononuclear cells.
Implementation Method 3
In some embodiments, isolating of the Pgp-positive cells from the sample is performed by any one or more methods selected from immunofluorescent methods, immunomagnetic methods, immunoaffinity methods, or combinations thereof.
Implementation Method 4
Pgp-positive cells were isolated by a number of techniques known in the art, such as magnetic activated cell sorting (MACS), in addition to conventional flow sorting.
Implementation Method 5
Pgp-positive cells were isolated by a number of techniques known in the art, such as magnetic activated cell sorting (MACS), in addition to conventional flow sorting.
Data Source
AI summary
Described herein are methods for selecting lymphocytes for adoptive cell therapy based on P-glycoprotein expression and compositions comprising same.