Coiled Coil Proteins with Polyarginine Tails
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Solution Overview
Problem
Current antigens with coiled coil domains lack effective modifications to enhance immunogenicity and binding properties to negatively charged polymers and nucleic acids, leading to suboptimal immune responses and potential adverse effects from nucleic acid signaling.
Innovation Solution
Modification of coiled coil proteins by linking short positively charged peptides, such as polyarginine sequences, to improve binding to negatively charged polymers like heparin and nucleic acids, and to enhance immunogenicity through genetic engineering or peptide synthesis, creating recombinant proteins with improved chromatographic properties and immune stimulation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If coiled coil proteins are modified by linking positively charged peptides, then binding to negatively charged polymers and nucleic acids is improved, but device complexity increases
Solution Approach 1:
The patent creates composite proteins by fusing coiled coil domains with positively charged peptide sequences (such as polyarginine motifs). This composite structure combines the oligomerization capability of coiled coils with the electrostatic binding capability of charged peptides, enabling simultaneous achievement of stable oligomeric structure and enhanced binding to negatively charged polymers and nucleic acids.
Solution Approach 2:
The modification introduces localized positively charged regions at specific positions (N-terminus, C-terminus, or internal positions) of the coiled coil protein without altering the entire protein structure. This localized modification approach maintains the overall coiled coil structure while adding specific binding functionality where needed, thus improving binding capacity without excessive complexity increase.
2Reliability
If coiled coil proteins are modified by linking positively charged peptides, then immunogenicity is improved, but manufacturing complexity increases
Solution Approach 1:
The fusion of coiled coil domains with positively charged peptide sequences creates recombinant proteins that can be produced using standard molecular cloning and expression systems. The modular nature of the fusion constructs allows for straightforward genetic engineering and scalable production, maintaining ease of manufacture while achieving enhanced immunogenicity.
Solution Approach 2:
The patent modifies specific parameters of the protein (adding charged peptide sequences) to enhance immunogenicity while maintaining production feasibility. The modifications are designed to be compatible with standard recombinant protein expression systems, allowing for straightforward optimization of expression conditions without requiring complex manufacturing processes.
3Ease of manufacture
If polyarginine tails are used to improve binding properties, then purification ease is improved, but protease susceptibility increases
Solution Approach 1:
The patent combines polyarginine motifs with coiled coil domains to create hybrid proteins that maintain the purification benefits of charged tags while gaining enhanced stability. The coiled coil portion provides structural stability and resistance to proteolytic degradation, while the charged peptide portion facilitates purification through ion-exchange or affinity chromatography.
Solution Approach 2:
The coiled coil domain acts as an intermediary protective element that shields the polyarginine tail from protease access. The stable oligomeric structure formed by coiled coils creates a protective environment for the charged peptide sequences, reducing their exposure to proteolytic enzymes while maintaining their charge-based purification properties.
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 modified proteins demonstrate enhanced binding to chromatography columns and increased immunogenicity, leading to stronger immune responses and reduced adverse effects by tightly binding nucleic acid ligands, thereby improving vaccine efficacy and safety.
Implementation Method 1
linking short positively charged peptides, such as polyarginine sequences, to improve binding to negatively charged polymers like heparin and nucleic acids
Data Source
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AI summary
The present application is related to a modified protein comprising a protein having a coiled coil domain and a peptide having the sequence such as shown in SEQ ID NO 1: ZXBBBBZ that is linked to the coiled coil domain wherein: • Z is any amino acid or is absent; • X is any amino acid; • B is an arginine (R) or a lysine (K). Said modified protein is in particular an antigen or a carrier protein, associated to an antigen. This modified protein has an increased affinity for negatively charged polymers such as nucleic acids or heparin, and shows an increased immunogenicity.