pMHC Multiplexer for High-Throughput T Cell Screening
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Solution Overview
Problem
Current methods for screening antigen-specific T cells are limited by low throughput, allowing only a few specificities to be tested in parallel, which hampers the development of personalized vaccines for cancer and infectious diseases.
Innovation Solution
The development of pMHC Multiplexers, which are spatially confined compositions of encoding molecules and peptides, enabling the simultaneous presentation of multiple pMHC complexes to T cells, facilitating higher-throughput screening and selection of antigen-specific T cells through fluorescence labeling and flow cytometry.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional fluorescent-labelled MHC Multimers are used for detection, then specific antigen-specific T cells can be detected, but only a few different antigen-specific T cell specificities can be tested in parallel
Solution Approach 1:
The patent combines multiple pMHC complexes with different antigen specificities into a single multiplexer structure. Each multiplexer contains multiple pMHC complexes that can simultaneously bind to different T cell specificities, allowing parallel detection of multiple antigen-specific T cell populations in a single assay. This merging approach directly increases both the throughput and versatility of the screening method.
Solution Approach 2:
The multiplexer structure serves multiple functions simultaneously: it presents multiple different antigen-specific pMHC complexes to T cells, enables detection of multiple T cell specificities in one experiment, and maintains the ability to identify specific pMHC-T cell receptor pairs. This multi-functionality resolves the contradiction by allowing a single reagent to perform what would traditionally require multiple separate assays.
2Productivity
If multiple pMHC complexes are presented simultaneously to T cells, then higher-throughput screening is enabled, but the complexity of the multiplexer structure increases
Solution Approach 1:
The multiplexer is segmented into discrete, modular components: multiple pMHC complexes are independently formed and then assembled into the multiplexer structure. This segmentation allows for standardized production of individual pMHC units that can be systematically combined, reducing the overall complexity of manufacturing and characterizing the multiplexer while maintaining the ability to present multiple antigens simultaneously.
Solution Approach 2:
The multiplexer employs a nested structure where multiple pMHC complexes are contained within a single multiplexer entity. The pMHC complexes are nested inside the multiplexer framework, allowing them to be presented together in a compact, organized manner. This nesting approach simplifies the overall structure by hierarchical organization, making the multiplexer easier to manufacture and characterize while maintaining high throughput capability.
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 approach allows for the identification and amplification of specific T cell clones, enabling the design of targeted vaccines and immunotherapies by enriching for antigen-specific T cells and revealing specific pMHC complex-T cell receptor pairs.
Implementation Method 1
A pMHC Multiplexer is a spatially confined composition of at least two molecules, an encoding molecule and a peptide (and/or protein) that is encoded by said encoding molecule
Implementation Method 2
Fluorescent-labelled MHC Multimers, consisting of multiple copies of a unique pMHC complex and carrying a fluorochrome compatible with flow cytometry
Implementation Method 3
T-lymphocytes (T cells) specifically recognize and bind target cells by interaction of their T cell receptors with Major Histocompatibility Complexes (MHC) on the target cells
Data Source
AI summary
This invention describes the production and properties of a pMHC Multiplexer. The pMHC Multiplexer is a spatially limited composition of two different molecules, an encoding molecule (i.e. an RNA or DNA molecule), and an encoded peptide, where said encoded peptide is encoded by said encoding molecule. Furthermore, the peptide is complexed to a MHC complex and thus is part of a pMHC complex. A preferred embodiment of the invention describes the production and properties of an example pMHC Multiplexer that is a phage particle carrying on its surface a number of identical pMHC complexes, where the peptide of the pMHC complexes is encoded by the DNA contained within the phage particle, and where a covalent or non-covalent bond links a phage coat protein with a pMHC complex and/or a pMHC Multimer. Another preferred embodiment of the invention describes the production and properties of an example pMHC Multiplexer that is a eukaryotic cell carrying on its surface a number of identical pMHC complexes, where the peptide of the pMHC complexes is encoded by the DNA contained within the cell. Yet another preferred embodiment of the invention describes the production and properties of an example pMHC Multiplexer where the encoding molecule is a DNA or RNA, and where the binding of pMHC Multiplexer to T cell receptor (TCR) can be detected by PCR-based analysis. Yet another preferred embodiment of the invention describes the production and properties of an example pMHC Multiplexer that comprises one or more identical pMHC complexes, where the encoding molecule is directly linked to at least one peptide (p) of one of the pMHC complexes, and thus, the peptide (p) of the pMHC complex(es) is encoded by said encoding molecule directly linked to it.