High-Affinity TCR Production via Bacteriophage Display
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Solution Overview
Problem
Current methods for generating T cell receptors (TCRs) often result in low-affinity TCRs due to central and peripheral T cell tolerance, making it difficult to recognize tumor cells expressing self-antigens, and existing techniques for enhancing affinity, such as bacteriophage display and amino acid substitution, are limited in generating high-affinity TCRs for tumor-associated antigens.
Innovation Solution
A method involving the transduction of cell populations with nucleic acids encoding TCR chains, followed by selection and isolation of cells expressing high-affinity TCRs specific for peptides of interest, using a bait and prey nucleic acid approach to enhance avidity and affinity, allowing for the production of recombinant cells capable of recognizing tumor cells with increased specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If TCRs are cloned from peripheral T cells, then the TCRs can be obtained through natural selection, but the affinity is low due to central and peripheral T cell tolerance
Solution Approach 1:
The patent introduces bacteriophage display technology to mutate and select TCR variants with enhanced affinity parameters. Through systematic random mutagenesis and selection processes, TCRs with affinities up to 1 million fold higher than wild-type can be obtained, directly addressing the low affinity problem while maintaining natural selection principles
Solution Approach 2:
The patent performs preliminary TCR cloning and characterization from peripheral T cells before applying affinity enhancement techniques. This preliminary action allows identification of starting TCRs that can then be optimized through bacteriophage display and amino acid substitution to achieve high affinity while understanding their natural recognition properties
2Measurement precision
If bacteriophage display mutation and selection technology is used to enhance TCR affinity, then high affinity TCRs can be generated, but the process complexity increases
Solution Approach 1:
The patent segments the TCR enhancement process into distinct modules: initial TCR cloning from peripheral T cells, bacteriophage display mutagenesis and selection, and final TCR variant characterization. This segmentation allows each step to be optimized independently and facilitates the integration of multiple techniques to achieve high affinity while managing overall process complexity
Solution Approach 2:
The patent uses bacteriophage display technology as an intermediary system to bridge natural TCR selection and high affinity TCR generation. The phage display platform serves as a mediator that allows in vitro selection and mutagenesis of TCR variants without requiring complex cellular manipulation, simplifying the overall process while achieving high affinity results
3Measurement precision
If amino acid substitution in TCR CDRs is performed to increase affinity, then TCR variants with higher affinity can be obtained, but the time and resources required increase
Solution Approach 1:
The patent performs preliminary identification of TCR CDR regions and their importance for antigen recognition before applying amino acid substitution. By pre-characterizing the TCR structure and antigen-binding properties, the substitution process can be targeted more efficiently, reducing the time and resources needed to achieve high affinity variants
Solution Approach 2:
The patent systematically varies amino acid parameters in the CDR regions through directed mutagenesis and selection. By changing specific parameters (amino acid composition, charge, hydrophobicity) in the CDR loops, high affinity TCR variants can be generated more rapidly than through random screening, reducing development time while achieving the desired affinity enhancement
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 enables the generation of high-affinity TCRs that can effectively recognize and target tumor cells, demonstrating increased avidity and affinity compared to native TCRs, potentially leading to improved clinical responses in cancer treatment.
Implementation Method 1
transducing a cell population with a nucleic acid which encodes either one of two polypeptide chains constituting a TCR
Implementation Method 2
generating TIL clones directly from melanoma patient tumor digests reveals a diversity of MART-1 reactive T cells with varying cellular avidities
Data Source
AI summary
Provided is a method for determining a TCR polypeptide chain that can form a TCR specific for a peptide of interest. Also provided are methods and compositions for producing a cell expressing a T cell receptor (TCR) specific for a peptide of interest, methods and compositions for producing a TCR chain nucleic acid and/or pair of TCR chain polypeptides and/or nucleic acids encoding a TCR, a cell population comprising the cell harboring the nucleic acids encoding a TCR obtained by said method, and a method for treating a disorder comprising administering to the subject said cell population.