Natural TCR Library Construction for Functional Antigen Binding
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Solution Overview
Problem
Existing methods for constructing T cell receptor (TCR) libraries face challenges in identifying functional TCRs with antigen binding specificity due to the complexity of TCR stability and folding issues, limited diversity in CDR3 sequences, and the absence of thymic selection, leading to non-functional or weakly binding TCRs.
Innovation Solution
A library of TCRs comprising alpha and beta chain variable domains derived from naturally occurring sequences, expressed from donor mRNA, without modifications or mutations, allowing for a more reliable identification of functional TCRs through natural pairing and thymic selection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If TCR libraries are created using existing methods with non-natural mutations, then diversity of TCR sequences is increased, but functionality and specificity of TCRs decrease
Solution Approach 1:
The patent changes the fundamental parameter of TCR library construction from introducing non-natural mutations to using natural TCR sequences directly from the immun repertoire. This parameter change resolves the contradiction by maintaining sequence diversity through natural variation while ensuring functionality and specificity through thymic selection history.
Solution Approach 2:
The patent copies natural TCR sequences from the immun repertoire rather than creating synthetic variants. This copying approach preserves the proven functionality and specificity of naturally selected TCRs while maintaining diversity through the inherent variation present in natural repertoires.
2Reliability
If TCR libraries retain natural diversity in CDR3 sequences, then antigen recognition capability is improved, but complexity of library construction increases
Solution Approach 1:
The patent extracts only the essential variable regions (Vα and Vβ domains) from natural TCR sequences, leaving behind the complex recombination and diversity generation processes. This extraction simplifies library construction while preserving the antigen recognition capability embedded in the natural CDR3 sequences.
Solution Approach 2:
The patent performs preliminary selection in the thymus where natural TCRs are already selected for functionality and specificity before being captured in the immun repertoire. This preliminary action eliminates the need for complex in vitro selection processes, simplifying library construction while ensuring antigen recognition capability.
3Reliability
If TCRs are isolated from natural repertoire, then specificity for antigen is improved, but difficulty in isolation and identification increases
Solution Approach 1:
The patent uses the immun repertoire itself as an intermediary source, capturing TCR sequences that have already been naturally selected for antigen specificity. This intermediary approach simplifies identification by providing pre-selected sequences that can be directly cloned and tested, reducing the difficulty of isolation and identification while maintaining high specificity.
4Productivity
If non-natural mutations are introduced in TCR libraries, then affinity maturation is accelerated, but loss of natural folding and stability increases
Solution Approach 1:
The patent performs preliminary optimization in the thymus where natural TCRs are selected for proper folding, stability, and functionality. By starting with pre-optimized natural sequences rather than introducing mutations de novo, the patent maintains stability while still allowing for subsequent affinity maturation through natural selection processes.
Data Source
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AI summary
The present invention relates to a library of particles, the library displaying a plurality of different T cell receptors (TCRs), wherein the plurality of TCRs consists essentially of TCRs comprising an alpha chain variable domain from a natural repertoire and a beta chain variable domain from a natural repertoire, wherein the alpha chain variable domain comprises a TRAV12-2 or a TRAV21 gene product and the beta chain variable domain comprises a TRBV6 gene product.