CTLA4-Ig Fusion Protein Formulation for Aggregation Control
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Fusion proteins, particularly CTLA4-Ig, are prone to instability due to aggregation, fragmentation, and oxidation, which affects their bioactivity and stability under various environmental conditions, necessitating the development of a suitable formulation that maintains stability and activity.
Innovation Solution
A stable pharmaceutical formulation of CTLA4-Ig fusion protein comprising phosphate buffer, sucrose or mannitol, histidine, and poloxamer, with histidine and sugar added during the tangential flow filtration process, enhancing colloidal stability and reducing aggregation and fragmentation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If fusion protein is formulated in solution, then it can be administered and used therapeutically, but it undergoes aggregation, deamidation, and oxidation leading to instability
Solution Approach 1:
The patent modifies formulation parameters including pH (maintained at 6.0-8.0 using phosphate buffer), temperature (storage at controlled conditions), and excipient concentrations to stabilize the fusion protein in solution, preventing aggregation and degradation while maintaining therapeutic usability
Solution Approach 2:
The patent introduces intermediary substances (excipients) such as sugars, amino acids, and surfactants that mediate between the fusion protein and the aqueous environment, protecting the protein from harmful interactions with water and stabilizing its structure during storage and administration
2Quantity of substance
If fusion protein concentration is increased to improve therapeutic dosage, then treatment effectiveness increases, but aggregation rate increases rapidly
Solution Approach 1:
The patent employs excipients as intermediary agents that interfere with protein-protein interactions, preventing aggregation even at high concentrations. These intermediaries create a protective environment that allows high protein concentration without loss of colloidal stability
Solution Approach 2:
The patent optimizes multiple formulation parameters simultaneously including pH, ionic strength, and excipient concentrations to expand the stable concentration range of the fusion protein, enabling high-dose formulations without aggregation
3Stability of the object's composition
If multiple excipients are added to stabilize the fusion protein, then protein stability improves, but formulation complexity increases
Solution Approach 1:
The patent combines multiple stabilizing functions into a coordinated excipient system where sugars provide structural stabilization, amino acids maintain pH and prevent deamidation, and surfactants prevent surface aggregation, achieving comprehensive protection through synergistic interaction rather than isolated components
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 formulation maintains the CTLA4-Ig fusion protein's stability for at least two weeks at 30°C with less than 10% in aggregate form, ensuring effective bioactivity and colloidal stability.
Implementation Method 1
comprising phosphate buffer at a pH ranging from pH 6.0 to pH 8.0
Implementation Method 2
sucrose or mannitol, histidine, and surfactant (poloxamer), wherein the fusion protein is a CTLA4-Ig molecule
Implementation Method 3
Deamidation can occur at aspargine or glutamine residues, resulting in a charge variant/s of the protein
Implementation Method 4
Oxidation of fusion proteins involves mainly methionine residues, and are generally influenced by external factors such as exposure to light and transition metal ions
Implementation Method 5
surfactant (poloxamer)
Data Source
AI summary
The present invention discloses a stable pharmaceutical formulation of a fusion protein, wherein the formulation contains buffer, sugar, amino acid and surfactant, and optionally includes salts. The disclosed fusion protein formulations are liquid formulations that are also suitable for lyophilization.