Bacillus subtilis Flagellin Variants for Stability and Reduced Immunogenicity
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Solution Overview
Problem
Existing flagellin proteins from Bacillus subtilis exhibit high immunogenicity, leading to side effects and reduced storage stability, limiting their clinical efficacy and utility in therapeutic applications.
Innovation Solution
Development of a Bacillus subtilis flagellin variant with an amino acid sequence having at least 50% identity to wild-type flagellin, featuring reduced immunogenicity and improved storage stability, achieved through specific amino acid substitutions, deletions, and modifications.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If wild-type flagellin is used, then immunogenicity is high (triggering strong immune response), but this leads to side effects and reduced storage stability
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of flagellin through specific substitutions (e.g., F194Y, F194W, L197F, L197Y, L197W mutations) to alter its immunogenic properties while maintaining TLR5 activation capability. This allows the protein to have reduced immunogenicity and improved storage stability without losing its physiological function
Solution Approach 2:
The patent converts the harmful high immunogenicity of wild-type flagellin into a benefit by creating variants that maintain sufficient immune activation (through TLR5 binding) while reducing excessive immune responses and side effects. The harmful property of strong immunogenicity is transformed into a controlled therapeutic benefit
2Reliability
If wild-type flagellin is used, then it activates TLR5 and provides therapeutic effects, but it causes side effects due to high immunogenicity
Solution Approach 1:
The patent modifies the flagellin amino acid sequence to change its immunogenic parameters while preserving TLR5 activation. Specific mutations (F194Y/W, L197F/Y/W) are introduced to reduce side effects while maintaining the therapeutic mechanism of TLR5-mediated immune activation
Solution Approach 2:
The patent applies local quality changes by making specific targeted amino acid substitutions at particular positions (194 and 197) in the flagellin sequence. These localized modifications affect immunogenicity and side effect profiles without disrupting the overall structure and function required for TLR5 activation and therapeutic efficacy
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 variant maintains physiological activity while significantly reducing immunogenicity and enhancing storage stability, offering potential therapeutic benefits in treating various conditions and serving as an effective vaccine adjuvant.
Implementation Method 1
TLR5 is an innate immune receptor located on the cell surface, consisting of an extracellular leucine-rich repeat (LRR) domain, a transmembrane domain, and an intracellular domain. TLR5 recognizes flagellin as a pathogen-associated molecular pattern using its extracellular domain and activates the MyD88-dependent signaling pathway and NF-κB-mediated inflammatory cytokine production.
Implementation Method 2
The patient's antibody response depends on the presence of B-cell epitopes and T-cell epitopes. When B-cell receptors in the body recognize and bind to administered foreign proteins, the foreign proteins are internalized into B-cells through receptor-mediated endocytosis and undergo protein hydrolysis. The resulting peptides are then presented by MHC class II molecules.
Data Source
AI summary
The present invention relates to a Bacillus subtilis flagellin variant and a use thereof, and more specifically to a flagellin variant and a use thereof, the flagellin variant having improved storage stability and reduced immunogenicity compared to flagellin derived from wild-type Bacillus subtilis.