Modified Coronavirus S Protein for Stable, High-Purity VLPs
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Solution Overview
Problem
Existing methods for producing coronavirus S protein face challenges in achieving high yield, homogeneity, and stability due to degradation in host cells and during purification, which hinders the efficient production of virus-like particles (VLPs) needed for widespread vaccination.
Innovation Solution
The introduction of specific amino acid modifications, such as N-glycosylation sites and deletions, stabilizes the coronavirus S protein, enhancing its integrity, resistance to degradation, and purity, thereby improving the production of VLPs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional methods are used to produce coronavirus S protein, then production process is simple, but the S protein exhibits degradation and low stability in host cells and during purification
Solution Approach 1:
The patent applies parameter changes by modifying specific amino acid residues in the S protein sequence (e.g., changing residue at position 24 from Aspartic acid to Asparagine, or deleting residues at positions 24-25) to alter the protein's stability parameters. These sequence modifications enable the S protein to resist degradation while maintaining its functional properties, directly resolving the stability issue without requiring complex production systems.
Solution Approach 2:
The patent extracts and removes specific problematic amino acid sequences (such as deleting residues at positions 24-25) that are responsible for degradation. By taking out these unstable segments, the remaining protein structure achieves improved stability and resistance to proteolytic cleavage, simplifying the overall production challenge rather than requiring complex stabilization systems.
2Productivity
If conventional S protein production methods are used, then manufacturing process is straightforward, but the yield of full-length S protein is low due to degradation
Solution Approach 1:
The patent applies preliminary anti-action by introducing amino acid modifications that preemptively prevent degradation before it occurs. By changing specific residues (e.g., D24N or del24-25) that are known degradation hotspots, the protein is pre-protected against proteolytic cleavage during host cell production and purification, thereby maintaining high yield of full-length S protein and reducing substance loss.
Solution Approach 2:
The patent implements beforehand cushioning by incorporating stabilizing amino acid changes that create a protective effect against degradation forces. These modifications (such as introducing Asparagine at position 24 or deleting positions 24-25) act as a buffer that absorbs or prevents the harmful effects of proteolytic enzymes, ensuring high yield of intact S protein throughout the production and purification process.
3Manufacturing precision
If conventional S protein production methods are used, then production process is simple, but the S protein lacks homogeneity and purity
Solution Approach 1:
The patent uses parameter changes in the amino acid sequence (specifically D24N or del24-25 modifications) to improve the homogeneity and purity of the S protein product. These changes create a more uniform protein structure that is resistant to degradation, resulting in a homogeneous product that requires simpler purification processes, thereby achieving high manufacturing precision without increasing system complexity.
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 S protein exhibits increased stability and purity, leading to higher yields of full-length S protein in VLPs, suitable for effective vaccination strategies.
Implementation Method 1
a substitution of one or more than one amino acid to introduce a N-glycosylation site at a position corresponding to positions 251, 252 or 253 of reference sequence SEQ ID NO: 1
Data Source
AI summary
The present disclosure relates to modified coronavirus S protein and virus-like particles (VLPs) comprising modified coronavirus S protein. The present invention also relates to methods of increasing the purity, and/or stability of coronavirus S protein or VLPs comprising modified coronavirus S protein in a host or host cell.