Barcoded MHC Complexes for High-Throughput TCR Identification
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Solution Overview
Problem
Current methods for identifying T cell receptors (TCRs) that bind to MHC:peptide complexes are inefficient and lack high-throughput capabilities, hindering the widespread adoption of adoptive T cell therapy (ACT) for cancer treatment.
Innovation Solution
A polypeptide comprising the amino acid sequence of a major histocompatibility complex (MHC) polypeptide, such as β2 microglobulin, is labeled with an identifier moiety, such as a nucleic acid moiety or HaloTag®, allowing for the identification of TCRs through trogocytosis and sortase-mediated interactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional TCR identification methods are used, then the process is simple, but the identification efficiency and throughput are low
Solution Approach 1:
The patent introduces MHC molecules as intermediary carriers that present peptide antigens to T cells. By labeling MHC molecules with unique identifiers, the system mediates the identification process between TCRs and peptides, enabling high-throughput screening without directly complexifying TCR analysis
Solution Approach 2:
The patent creates labeled copies of MHC:peptide complexes where the MHC molecule is tagged with unique identifiers (such as nucleic acid barcodes or fluorescent labels). These labeled copies serve as surrogates that carry identification information, allowing parallel processing and high-throughput identification of multiple TCR-specificities simultaneously
2Measurement precision
If MHC molecules are labeled with identifier moieties, then TCR identification precision is improved, but the complexity of the polypeptide structure increases
Solution Approach 1:
The patent segments the identification system by separating the functional MHC:peptide complex from the identification label. The MHC molecule retains its natural peptide-binding function while the identifier moiety (such as a nucleic acid barcode or fluorescent tag) is attached as a distinct modular component, allowing independent optimization of each function
Solution Approach 2:
The patent creates composite MHC molecules that combine the native MHC polypeptide with identifier moieties such as nucleic acid barcodes, fluorescent labels, or affinity tags. These composite structures integrate the antigen-presenting function of MHC with the detectable properties of the label, achieving both functional integrity and identification capability
3Speed
If high-throughput TCR identification is implemented, then the speed of TCR discovery increases, but the complexity of the experimental process increases
Solution Approach 1:
The patent performs preliminary labeling of MHC molecules with unique identifiers before the TCR identification experiment. This advance preparation creates a library of pre-labeled MHC:peptide complexes that can be directly used in high-throughput screening, eliminating the need for complex real-time labeling procedures during the actual identification process
Solution Approach 2:
The patent replaces complex mechanical or manual TCR identification procedures with molecular-level detection systems. By using nucleic acid barcodes that can be read by sequencing technologies or fluorescent labels detectable by flow cytometry, the system substitutes complex manual analysis with automated, high-throughput molecular detection methods
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
Enables rapid and precise identification of TCRs that bind to MHC:peptide complexes, facilitating the development of effective adoptive T cell therapies by enhancing the understanding of TCR-MHC interactions.
Implementation Method 1
a moiety facilitating labelling of the polypeptide with an identifier moiety, such as a nucleic acid moiety or HaloTag®
Implementation Method 2
Identification of HLA:peptide targets of orphan TCRs has been achieved by evaluation of trogocytosis of membrane contents from the T cell onto the HLA:peptide-presenting target cell
Implementation Method 3
Trogocytosis is a well-established process that occurs bidirectionally between T cells and cells expressing MHC:peptide complexes recognised by the TCRs they express, during immune interactions
Data Source
AI summary
The present disclosure relates to the fields of molecular biology and immunology.