Thermostable FGF10 Polypeptide Engineering for Longer Activity
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Solution Overview
Problem
The natural instability and low thermostability of Fibroblast Growth Factor 10 (FGF10) limit its practical use in medical and cosmetic applications due to rapid degradation and loss of biological activity, necessitating frequent replacement and potential toxicity from unfolded or aggregated protein forms.
Innovation Solution
Development of a thermostable engineered FGF10 polypeptide with specific amino acid substitutions, such as L152F, and optionally V123I, Q175E, and N181D, enhancing its thermal stability and maintaining biological activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If wild-type FGF10 is used, then biological activity is maintained, but thermal stability is poor and degradation occurs rapidly
Solution Approach 1:
The patent applies parameter changes by introducing specific amino acid substitutions (L152F, V123I, Q175E, N181D) into the FGF10 protein sequence. These substitutions modify the protein's molecular parameters to enhance thermal stability and reduce degradation, thereby extending its half-life while preserving biological activity. The L152F substitution alone increases melting temperature by 7-19°C, and combinations of multiple substitutions provide even greater stabilization.
2Reliability
If FGF10 is used in cell culture, then cell proliferation and differentiation are promoted, but protein degradation leads to loss of activity and potential toxicity
Solution Approach 1:
The patent converts the harmful effect of protein degradation into a benefit by strategically introducing amino acid substitutions that prevent degradation. The substitutions target regions prone to unfolding and aggregation, transforming the vulnerable wild-type protein into a thermostable variant that maintains its native conformation and biological activity without forming toxic aggregates.
Solution Approach 2:
The patent modifies the protein's structural parameters through amino acid substitutions (L152F, V123I, Q175E, N181D) to enhance folding stability and resistance to denaturation. These parameter changes ensure the protein remains in its functional conformation under cell culture conditions, preventing aggregation and toxicity while maintaining proliferative and differentiative effects.
3Reliability
If additional stabilizing compounds are added to maintain FGF10 activity, then protein stability improves, but formulation complexity increases
Solution Approach 1:
The patent enables the protein to serve itself by incorporating stabilizing amino acid substitutions directly into its sequence. The engineered FGF10 variants (with substitutions such as L152F, V123I, Q175E, N181D) inherently possess enhanced stability without requiring external stabilizing compounds. This self-service approach eliminates the need for complex formulations containing additives like heparin or other stabilizing agents.
Data Source
AI summary
The invention relates to a thermostable FGF10 polypeptide possessing FGF10 activity and having or comprising at least 85% sequence identity to SEQ ID NO:3 that has or comprises an amino acid sequence from Ser69 to Ser208 of SEQ ID NO:1, or to SEQ ID NO:5 that has or comprises an amino acid sequence from Leu40 to Ser208 of SEQ ID NO:1, or the fragments thereof, comprising at least an amino acid substitution L152F. Preferably, any of amino acid substitutions V123I, Q175E, and N181D can be further included. The invention further discloses the use of subjected thermostable FGF10 polypeptides in regenerative medicine or other related medical applications or cosmetics. Further, it discloses a culture medium comprising subjected thermostable FGF10 polypeptides suitable for proliferation and differentiation of the human embryonic stem cells or formation and differentiation of spheroids and organoids.