CAR Library Screening for Functional scFvs in CAR-T Cells
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for selecting single-chain antibodies (scFvs) capable of binding to target antigens do not ensure their functionality in chimeric antigen receptor-expressing T cells (CAR-T cells).
Innovation Solution
A CAR library is developed, comprising nucleic acids encoding chimeric antigen receptors (CARs) with specific antigen-binding domains, transmembrane domains, and intracellular signaling domains, where the scFv includes heavy- and light-chain variable regions meeting predefined conditions, allowing direct screening for functionality in CAR-T cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If hybridoma library or phage display technique is used to select scFv, then scFv binding capability to target antigen is achieved, but functionality in CAR-T cells cannot be ensured
Solution Approach 1:
The patent merges the scFv screening process with CAR-T cell functionality assessment by expressing scFvs in T cells and evaluating their functional activity directly. This integration ensures that selected scFvs are not only capable of binding target antigens but also functional when expressed in CAR-T cells, resolving the reliability issue without requiring separate validation steps.
Solution Approach 2:
The patent performs preliminary functional assessment during the scFv selection process itself. By expressing scFvs in T cells and evaluating their activity against target antigens before final selection, the method ensures functionality is built into the selection criteria rather than added as a subsequent validation step.
2Reliability
If scFv is expressed in T cells for functional examination, then functionality is confirmed, but time and resources are consumed
Solution Approach 1:
The patent performs functional assessment during the scFv selection process itself. By expressing scFvs in T cells and evaluating their activity against target antigens before final selection, the method ensures functionality is built into the selection criteria rather than added as a subsequent validation step.
Solution Approach 2:
The patent incorporates feedback from functional activity measurements directly into the scFv selection process. By monitoring T cell activity and scFv binding function simultaneously, the method uses real-time feedback to guide selection, eliminating the need for separate post-selection validation and reducing overall screening time.
Data Source
AI summary
Provided are a CAR library used to screen scFvs that can be functional in CAR-T cells, and an scFv manufacturing method in which the CAR library is used. A chimeric antigen receptor (CAR) library of the present invention includes nucleic acids coding for first CARs. Each of the first CARs includes a first antigen-binding domain, a first transmembrane domain, and a first intracellular signaling domain. The first antigen-binding domain includes a first single-chain antibody (scFv) to be screened for the ability to bind to a target antigen. The first scFv includes a first heavy-chain variable region and a first light-chain variable region. The first heavy-chain variable region and the first light-chain variable region meet a predetermined condition.