Albumin Binding Polypeptide with Arg-to-Lys Substitution for Protease Stability
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Solution Overview
Problem
Current polypeptides with high albumin affinity often lack resistance to enzymatic cleavage, particularly by clostripain, which limits their stability and effectiveness in biomedical and biotechnological applications.
Innovation Solution
Development of albumin binding polypeptides with a specific albumin binding motif, including an Arg-to-Lys substitution at position 8, which enhances resistance to clostripain cleavage and maintains high albumin affinity, integrated into three-helix bundle protein domains from bacterial receptor proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If polypeptides with high albumin affinity are designed using conventional albumin binding motifs, then albumin binding activity is improved, but resistance to enzymatic cleavage by clostripain deteriorates
Solution Approach 1:
The patent applies local quality by making a specific local modification (Arg-to-Lys substitution) at position 8 within the albumin binding motif, while keeping the rest of the motif sequence intact. This localized change specifically addresses clostripain cleavage susceptibility without disrupting the overall albumin binding capability of the motif.
Solution Approach 2:
The patent changes the amino acid parameter at position 8 from Arginine (Arg) to Lysine (Lys). This parameter change modifies the chemical properties of the polypeptide at the cleavage site, reducing susceptibility to clostripain while preserving albumin binding affinity, thus resolving the contradiction between binding activity and enzymatic stability.
2Stability of the object's composition
If Arg-to-Lys substitution is made at position 8 to increase resistance to clostripain cleavage, then protease stability is improved, but potential impact on albumin affinity must be considered
Solution Approach 1:
The patent applies parameter changes by substituting Arginine with Lysine at position 8, modifying the chemical properties (charge, side chain structure) at this specific location. This change increases protease stability while the patent demonstrates through experimentation that albumin affinity is maintained, thus resolving the potential trade-off between these two properties.
Solution Approach 2:
The modification is applied locally at position 8 rather than throughout the entire motif, allowing the overall structure and function to be preserved while achieving the desired increase in protease stability. The local change minimizes disruption to albumin binding while providing the stability benefit.
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 albumin binding polypeptides exhibit superior protease stability and albumin binding activity, with variants showing reduced cleavage by clostripain and maintaining strong affinity for albumin, thus extending their utility in therapeutic and biotechnological applications.
Implementation Method 1
Several strategies have been reported to either covalently couple proteins directly to serum albumins or to a peptide or protein that will allow in vivo association to serum albumins
Implementation Method 2
Current polypeptides with high albumin affinity often lack resistance to enzymatic cleavage, particularly by clostripain
Data Source
AI summary
The present disclosure relates to a class of engineered polypeptides having a binding affinity for albumin. In particular, the present invention relates to albumin binding polypeptides which have a high resistance to enzymatic cleavage. The disclosure provides an albumin binding polypeptide comprising an albumin binding motif, which motif consists of the amino acid sequence GVSDFYKKLI XaKAKTVEGVE ALKXbXcI (SEQ ID NO:29).