Rh5 Polypeptide Expression for Malaria Vaccine Cross-Protection
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Solution Overview
Problem
Current vaccines for malaria, particularly targeting the erythrocytic stage of Plasmodium falciparum, face challenges in producing functional recombinant parasite proteins due to high AT content, low complexity regions, and difficulties in correct folding and solubilization, leading to inadequate cross-protection across multiple strains.
Innovation Solution
A method for expressing polypeptides, including Rh5, using a eukaryotic expression system that prevents N-glycosylation, includes an exogenous signal sequence for secretion, and codon optimization, enabling the production of correctly folded and immunogenic proteins that can bind to CD147, a critical receptor for erythrocyte invasion, thereby blocking parasite entry across multiple strains.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If recombinant Plasmodium proteins are expressed in heterologous systems, then production scalability is improved, but correct folding and functional activity deteriorate due to inability to form disulfide bonds in reducing environments
Solution Approach 1:
The patent uses an oxidizing environment (peroxisome or endoplasmic reticulum) as an intermediary system that mediates between the need for scalable heterologous expression and the requirement for correct disulfide bond formation. The signal sequence directs proteins to these compartments where oxidative conditions enable proper folding while maintaining production scalability.
Solution Approach 2:
The patent changes the redox parameter of the expression environment by targeting proteins to oxidizing compartments (peroxisome or ER) instead of standard reducing cytoplasmic expression systems. This parameter change enables disulfide bond formation and correct protein folding while maintaining the benefits of heterologous expression.
2Adaptability or versatility
If multiple Plasmodium strains are targeted to achieve cross-protection, then vaccine efficacy across strains is improved, but vaccine complexity and development difficulty worsen
Solution Approach 1:
The patent identifies conserved regions within Plasmodium proteins that are universal across multiple strains. By targeting these conserved epitopes, a single vaccine composition can provide cross-protection against multiple strains, achieving versatility without increasing complexity.
Solution Approach 2:
The patent uses computational methods to identify and copy conserved sequence patterns across different Plasmodium strains. These copied conserved regions are then used as vaccine targets, allowing development of vaccines that work across strains without having to develop separate vaccines for each strain.
3Ease of operation
If membrane-tethered proteins are solubilized for vaccine production, then accessibility to immunogenic epitopes is improved, but protein stability and correct conformation worsen due to disruption of transmembrane domains
Solution Approach 1:
The patent extracts only the extracellular domain (ECD) of membrane-tethered proteins, separating it from the transmembrane and cytoplasmic domains. This extraction maintains the immunogenic epitopes in the ECD while avoiding the stability issues associated with solubilizing entire membrane proteins, including their hydrophobic transmembrane regions.
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 approach allows for the production of functional and immunogenic polypeptides that effectively inhibit Plasmodium infection by targeting the Rh5-CD147 interaction, providing cross-strain protection and potentially leading to a more effective malaria vaccine.
Implementation Method 1
the nucleic acid encodes an exogenous eukaryotic signal sequence effective to deliver the polypeptide into the secretory pathway of the eukaryotic cell
Implementation Method 2
An antibody which binds to CD147, for use in the prevention and/or treatment of Plasmodial infection and/or disease
Data Source
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AI summary
Immunogenic compositions and vaccines against Plasmodialinfection comprising an Rh polypeptide or a fragment or variant thereof are dislcosed. Also disclosed are Rh5 polypeptides or fragments or variants thereof capable of binding CD147 and conferring protection against infection and/or disease caused by multiple Plasmodial strains or Plasmodialspecies, inhibitors of the interaction between Rh5 and CD147 and methods for producing polypeptides in a mammalian expression system.