Stabilized Coronavirus Spike Glycoprotein for Freeze-Thaw and Lyophilization
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current SARS-CoV-2 spike proteins face challenges in stability during storage, manufacturing, freeze/thaw cycles, and lyophilization, which affect vaccine efficacy and immunogenicity, particularly against emerging variants.
Innovation Solution
The development of mutant spike proteins with modifications such as short flexible or rigid peptide linkers, additional disulfide bonds, and proline mutations to enhance trimeric stability, allowing for improved expression and stability, including temperature, freeze/thaw, and lyophilization resistance.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If the native SARS-CoV-2 spike protein structure is used, then the protein maintains natural antigenicity and immunogenicity, but the protein exhibits poor stability during storage, manufacturing, freeze/thaw cycles, and lyophilization
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the spike protein through specific mutations (e.g., proline substitutions at positions 986 and 987, deletions of the furin cleavage site). These parameter changes in the protein structure significantly improve stability during storage, freeze/thaw cycles, and lyophilization while preserving antigenicity and immunogenicity.
2Stability of the object's composition
If the native SARS-CoV-2 spike protein structure is used, then the protein retains native antigenicity, but the protein undergoes conformational changes from pre-fusion to post-fusion state affecting vaccine efficacy
Solution Approach 1:
The patent applies preliminary action by introducing stabilizing mutations (particularly proline substitutions) that pre-lock the spike protein in its pre-fusion conformation. This preliminary structural stabilization prevents unwanted conformational changes to the post-fusion state during storage and manufacturing, ensuring the protein maintains its immunogenic pre-fusion structure throughout the vaccine process.
3Strength
If additional disulfide bonds and proline mutations are introduced, then trimeric stability and thermostability are enhanced, but the protein structure becomes more complex
Solution Approach 1:
The patent applies local quality by introducing specific localized modifications (disulfide bonds at particular cysteine positions and proline mutations at specific residues) rather than global structural changes. These localized modifications strategically enhance trimeric stability and thermostability while minimizing overall structural complexity and maintaining the protein's fundamental architecture and antigenic properties.
Data Source
AI summary
Provided herein are mutant coronavirus spike proteins, methods of making and using, vaccines, vectors and nucleic acids, comprising at least one of the following modifications: a short flexible peptide linker or a rigid peptide linker in place of the furin cleavage site loop to genetically link an S1 and S2 subunit; at least one additional disulfide bond; or 1, 2, 3, 4, or 5 proline mutations for greater trimeric stability, wherein the mutant coronavirus spike protein has: a higher stability or a higher level of expression when compared to a non-modified coronavirus spike protein. Coronavirus is SARS, MERS, 229E (alpha), NL63 (alpha), OC43 (beta), HKU1 (beta), SARS-CoV-2, or an emerging variant thereof. Cunent SARS-CoV-2 variants include, e.g., B.1.1.7, B.1.1.7 with E484K, B.1.135, B.1.351, P.1, B.1.427, D614G, B.1.1351, or B.1.429, Lambda (i.e., C.37), Mu (i.e., B.1.621), Omicron (B.1.1.529) or a variant (including but not limited to BA.1, BA.2, or BA.3) thereof, and others.