Stabilizing MHC Class II Heterodimers via Disulphide Bonds
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Solution Overview
Problem
The production of stable, fully functional soluble MHC class II molecules is hindered by their intrinsic instability and low production levels, making it challenging to develop recombinant forms for immunomodulator screening and therapeutic applications.
Innovation Solution
The introduction of an artificial disulphide bond between the α2 and β2 domains of the MHC class II molecule, using engineered cysteine residues, stabilizes the heterodimer and allows for its production in a prokaryotic expression system, enhancing stability and functionality.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If soluble MHC class II molecules are produced using conventional methods, then production levels are low and molecular stability is poor, but introducing artificial disulphide bonds increases stability and enables prokaryotic expression
Solution Approach 1:
The patent introduces artificial disulphide bonds between the α2 and β2 domains of MHC class II molecules, fundamentally changing the molecular structure parameter. This structural modification increases stability and enables prokaryotic expression, directly resolving the contradiction between molecular stability and production ease.
Solution Approach 2:
The invention creates a composite structure by combining natural MHC class II heterodimer components with artificial disulphide bond linkages. This composite approach integrates the functional elements of native MHC molecules while adding stabilizing covalent bonds that enable soluble production in prokaryotic systems.
2Productivity
If conventional production methods are used for MHC class II molecules, then production levels are low, but using prokaryotic expression systems with disulphide bond stabilization significantly increases production capability
Solution Approach 1:
The patent modifies the molecular structure by introducing artificial disulphide bonds, which changes the production parameters to enable prokaryotic expression. This structural parameter change allows high-level production in bacteria while maintaining molecular reliability through the stabilizing disulphide linkages.
3Adaptability or versatility
If native MHC class II molecules are produced, then they maintain natural structure, but they lack stability and cannot be produced in soluble form at high levels
Solution Approach 1:
The invention changes the structural parameters of MHC class II molecules by introducing artificial disulphide bonds between α2 and β2 domains. This parameter change enables soluble production capability while simultaneously improving molecular stability, allowing high-level production in prokaryotic systems.
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 results in stable, functional MHC class II molecules that can specifically bind peptides and interact with T cell receptors, enabling effective T cell staining and immunomodulator screening, overcoming the limitations of previous methods.
Implementation Method 1
The introduction of an artificial disulphide bond between the α2 and β2 domains of the MHC class II molecule, using engineered cysteine residues, stabilizes the heterodimer
Data Source
Figure 1A~1C
Figure 2
Figure 3A~3B
AI summary
The present invention relates to disulphide bond stabilized recombinant MHC class II molecules. In particular, the present invention provides a recombinant MHC class II molecule, which comprises: (i) all or part of the extracellular portion of an MHC class II α chain; (ii) all or part of the extracellular portion of an MHC class II β chain; wherein (i) and (ii) provide a functional peptide binding domain and wherein (i) and (ii) are linked by a disulphide bond between cysteine residues located in the α2 domain of said α chain and the β2 domain of said β chain, wherein said cysteine residues are not present in native MHC class II α2 and β2 domains. Methods of producing these molecules in prokaryotic systems and various uses of these molecules form further aspects.