Single Variable Domain Formulation to Limit Stress-Induced Dimerization
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Solution Overview
Problem
Existing technologies face challenges in stabilizing and maintaining the monomeric form of single variable domains, leading to dimer formation that can affect binding functionality and stability, particularly under stress conditions such as high concentration and temperature.
Innovation Solution
Formulations containing excipients like polyols and non-reducing sugars, such as mannitol, sorbitol, and trehalose, are used to stabilize single variable domains, reducing dimer formation and maintaining binding activity under stress conditions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If single variable domains are stored under stress conditions (high concentration and temperature), then binding activity may be maintained, but dimer formation increases reducing stability
Solution Approach 1:
The patent introduces excipients (polyols, non-reducing sugars, amino acids) as intermediary substances that mediate between the single variable domains and the stress conditions. These excipients form protective complexes with the protein domains, preventing direct harmful interactions that lead to dimerization while allowing the domains to maintain their binding function under stress conditions.
Solution Approach 2:
The patent modifies the physical and chemical parameters of the storage environment by adjusting excipient concentrations, pH levels, and ionic strength. These parameter changes create optimal conditions that favor monomeric stability while preserving binding activity, effectively resolving the contradiction between maintaining reliability under stress and preventing compositional instability.
2Stability of the object's composition
If excipients are added to suppress dimer formation, then monomeric stability improves, but formulation complexity increases
Solution Approach 1:
The patent applies local quality by selecting specific excipients with particular properties (polyols for hydrophilic protection, non-reducing sugars for structural stabilization, amino acids for charge interaction) and positioning them at optimal concentrations in the formulation. Each excipient class addresses specific local stability issues in the formulation, allowing complex functionality through targeted local improvements rather than uniform approaches.
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 formulations achieve high stability and retention of binding activity, with less than 10% dimer formation and at least 80% activity retention after storage for up to two years under stress conditions, enhancing the stability and efficacy of single variable domain polypeptides.
Implementation Method 1
adding excipients that increase the melting temperature of the single variable domain such as for example by adding mannitol, other polyols or reducing sugars to a liquid formulation
Data Source
AI summary
In a broad aspect the present invention generally relates to novel dimer-complexes (herein called “non-fused-dimers” or NFDs) comprising single variable domains, methods of making these complexes and uses thereof. These non-covalently bound dimer-complexes consist of two identical monomers that each comprises of one or more single variable domains (homodimers) or of two different monomers that each comprises on or more single variable domains (heterodimers). The subject NFDs have typically altered e.g. improved binding characteristics over their monomeric counterpart. The NFDs of the invention may further be engineered through linkage by a flexible peptide or cysteines in order to improve the stability. This invention also describes conditions under which such NFDs are formed and conditions under which the formation of such dimers can be avoided.