Peptide Tag Binding Partner Isopeptide Bond Formation
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Solution Overview
Problem
Current peptide tag and binding partner systems for forming isopeptide bonds lack stability and efficiency in biological conditions, leading to rapid dissociation and limited protease resistance.
Innovation Solution
Designing peptide tag and binding partner pairs that can spontaneously form isopeptide bonds, where the peptide tag and binding partner are expressed as separate fragments, allowing for covalent reconstitution and enhanced stability under biological conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If peptide tags and binding partners are used to form isopeptide bonds, then binding stability and protease resistance are improved, but the interaction efficiency and binding efficacy are insufficient under biological conditions
Solution Approach 1:
The invention divides the original protein into separate peptide tag and binding partner fragments, each containing specific residues required for isopeptide bond formation. This segmentation allows the fragments to be independently optimized and expressed, then reconstituted through spontaneous isopeptide bond formation to achieve stable and reliable interactions under biological conditions.
Solution Approach 2:
The invention optimizes specific parameters of the peptide tag and binding partner sequences, including the selection of residues involved in isopeptide bond formation (such as lysine, asparagine, aspartic acid, glutamine, or glutamic acid residues), to enhance both binding stability and interaction efficiency. By carefully selecting and positioning these residues, the system achieves high reliability while maintaining stability.
2Duration of action of stationary object
If peptide tags and binding partners interact via isopeptide bonds, then irreversible binding and protease resistance are achieved, but the spontaneous formation efficiency is limited
Solution Approach 1:
The invention incorporates all necessary residues for isopeptide bond formation within the peptide tag and binding partner sequences themselves before the interaction occurs. This preliminary inclusion of reactive residues (such as lysine, asparagine, aspartic acid, glutamine, or glutamic acid) enables spontaneous and efficient bond formation upon encounter, eliminating the need for external enzymes or catalysts and achieving both irreversibility and high productivity.
3Ease of manufacture
If separate fragments are used for peptide tag and binding partner, then covalent reconstitution is enabled, but the complexity of designing and identifying suitable pairs increases
Solution Approach 1:
The invention focuses on optimizing specific local regions of the protein sequence that contain the residues necessary for isopeptide bond formation. By concentrating the design effort on these critical local areas (such as positioning lysine, asparagine, aspartic acid, glutamine, or glutamic acid residues at specific positions), rather than optimizing the entire sequence, the design complexity is reduced while still enabling effective covalent reconstitution of separate fragments.
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 solution provides improved irreversible interactions and increased binding efficacy and stability, enabling effective protein manipulation and analysis, particularly in applications like cancer immunotherapy and vaccine development.
Implementation Method 1
peptide tags and binding partners which are capable of interacting via the spontaneous formation of an isopeptide bond
Data Source
AI summary
The present invention relates to peptide tags and binding partners which are capable of interacting via the spontaneous formation of an isopeptide bond, as well as to associated peptide pairs and methods for designing peptide tags, binding partners and peptide pairs with improved properties.