Stabilized MHC Covalent Bridges for High-Throughput TCR Screening
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Solution Overview
Problem
Current methods for high-throughput screening of TCR-binding peptide ligand/MHC molecule complexes face challenges in specificity and safety due to off-target toxicities and the difficulty in generating high-quality pMHC complexes, especially for clinical settings without advanced facilities.
Innovation Solution
A method involving stabilized MHC molecules with artificially introduced covalent bridges between specific amino acids, allowing for the formation of peptide ligand/MHC complexes that are stable, recognizable by TCRs with high specificity and selectivity, and suitable for high-throughput screening.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional MHC molecules are used for high-throughput screening, then the screening can be performed, but the complexes lack stability and specificity leading to off-target toxicities
Solution Approach 1:
The patent modifies the MHC molecule structure by introducing covalent bridges between specific amino acid pairs (e.g., Cys84-Cys139, Cys84-Cys85, or Cys139-Cys84/85) to change the structural parameters of the MHC molecule. This stabilization enhances the specificity of pMHC-TCR interactions and reduces off-target toxicities by ensuring proper folding and conformational integrity of the MHC molecule during high-throughput screening.
2Manufacturing precision
If advanced facilities are used for generating high-quality pMHC complexes, then the quality improves, but the method becomes inaccessible for clinical settings without advanced facilities
Solution Approach 1:
The patent extracts the essential function of advanced facility-based MHC stabilization by incorporating covalent bridges directly into the MHC molecule structure itself. This allows clinical settings without advanced facilities to generate high-quality pMHC complexes using simpler procedures, as the stabilized MHC molecules can be produced through standard recombinant protein expression followed by peptide loading, eliminating the need for complex facility-based stabilization methods.
3Stability of the object's composition
If the MHC molecule structure is stabilized with covalent bridges, then the complex stability and affinity improve, but the structural complexity increases
Solution Approach 1:
The patent applies covalent bridges at specific localized positions within the MHC molecule structure (such as between Cys84 and Cys139, or Cys84 and Cys85, or Cys139 and Cys84/85) rather than throughout the entire molecule. This localized stabilization approach enhances complex stability and affinity while minimizing the increase in overall structural complexity, as only specific regions require modification rather than global structural changes.
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
The method enables the generation of stable, high-affinity pMHC complexes that are representative of wild-type variants, facilitating accurate affinity measurements and identifying cross-reactive peptides, thus enhancing the safety and efficacy of therapeutic candidates.
Implementation Method 1
MHC molecule comprises at least one artificially introduced covalent bridge between amino acids of the alpha1 domain and amino acids of the alpha2 domain
Implementation Method 2
contacting said suitably stabilized MHC molecule with a multitude of peptide ligands thereof, to form peptide ligand/MHC (pMHC) molecule complexes
Implementation Method 3
screening said pMHC molecule complexes for TCR-binding
Data Source
AI summary
The present invention relates to a method for high throughput screening for a TCR-binding peptide ligand/MHC molecule complex, comprising a stabilized peptide-MHC molecule and respective uses of said method. The present invention further relates to polypeptides comprising or consisting of stabilized MHC molecules or peptide binding fragments thereof, pharmaceutical compositions comprising said polypeptides, vaccines comprising said pharmaceutical composition and uses of said vaccine for the manufacturing of a medicament and/or in the prevention of cancer The present invention further relates to nucleic acids encoding said polypeptides and vectors comprising said nucleic acids.