CMV Virus-Like Particle Charge Modification for Aggregation Resistance
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Solution Overview
Problem
Existing virus-like particle (VLP) based vaccines face challenges in stability, aggregation, and manufacturability, which are critical for clinical trials and market launch, particularly under varying conditions and high salt solutions, impacting their processability and purification.
Innovation Solution
Incorporating stretches of consecutive negative amino acids, such as glutamic acid and aspartic acid, into the βB-βC loop of cucumber mosaic virus (CMV) polypeptides enhances the stability and prevents aggregation of modified VLPs, allowing for improved processability and purification through ion-exchange chromatography.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional CMV VLPs are used for antigen presentation, then immunogenicity is achieved, but stability and aggregation control deteriorate under varying conditions and high salt solutions
Solution Approach 1:
The patent modifies the amino acid sequence of the CMV coat protein by introducing stretches of consecutive negative amino acids (glutamic acid and/or aspartic acid) at specific positions (βB-βC loop region). This parameter change in the protein composition alters the surface charge characteristics of the VLPs, enhancing their stability and preventing aggregation under varying ionic strengths and temperatures while preserving immunogenicity.
2Ease of manufacture
If chemical linker coupling technology is used to present antigens on VLP surface, then antigen presentation capability is improved, but manufacturability and purification difficulty increase
Solution Approach 1:
The modified CMV VLPs with enhanced surface charge properties exhibit improved self-purification characteristics through ion-exchange chromatography. The consistent negative surface charge allows for efficient separation and purification without requiring complex chemical linker coupling technologies, simplifying the manufacturing process while maintaining antigen presentation capability.
3Stability of the object's composition
If additional negative charge is introduced in coat protein to improve stability, then resistance to aggregation is improved, but VLP formation capability may deteriorate
Solution Approach 1:
The patent introduces stretches of consecutive negative amino acids at specific local regions (βB-βC loop) of the coat protein rather than uniformly throughout the entire protein sequence. This localized modification approach enhances surface charge and aggregation resistance at the VLP surface while preserving the overall structural integrity and self-assembly capability of the coat protein necessary for VLP formation.
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 CMV VLPs maintain structural integrity and immunogenicity, reducing aggregation and enabling scalable manufacturing, thus meeting the requirements for drug development and product registration.
Implementation Method 1
the specific insertion of these stretches of consecutive negative amino acids selected from glutamic acid and aspartic acid into the CMV polypeptides and coat proteins unexpectedly led even to improvements in stability of the resulting modified CMV VLPs
Implementation Method 2
the improved stability in higher salt solution arising from the surface charge modifications to the CMV VLPs is highly beneficial or even essential for its processability and purification by ion-exchange chromatography
Data Source
AI summary
The present invention relates to a modified virus-like particle (VLP) of cucumber mosaic virus (CMV) comprising at least one chimeric CMV polypeptide, wherein said at least one chimeric CMV polypeptide comprises, preferably consists of (i) a CMV polypeptide, wherein said CMV polypeptide comprises a coat protein of CMV or an amino acid sequence having a sequence identity of at least 75% with SEQ ID NO:48; and (ii) a polypeptide comprising, preferably consisting of, a stretch of consecutive negative amino acids, wherein said negative amino acids are independently selected from aspartic acid or glutamic acid, wherein said polypeptide is inserted between any amino acid residue of said CMV polypeptide corresponding to any amino acid residue between position 75 and position 85 of SEQ ID NO:48, as well as to compositions and pharmaceutical compositions comprising such modified VLPs to which antigens are linked, which compositions preferably serve as vaccine platform for generating immune responses, in particular antibody responses, against said antigens linked to the modified CMV VLPs.