AAV Vector Formulations for Lyophilization Stability
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Solution Overview
Problem
Existing recombinant adeno-associated virus (rAAV) formulations lack stability during manufacture, shipping, storage, and administration, necessitating improved formulations that maintain product quality under various conditions.
Innovation Solution
A formulation comprising recombinant adeno-associated virus (rAAV) particles with Tris as a buffering agent, a sugar such as sucrose, and an amorphous salt like sodium citrate, optionally with a non-ionic surfactant like poloxamer 188, is developed for lyophilization, which enhances stability and reduces genome release.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional rAAV formulations are used, then the product can be manufactured and shipped, but the formulation lacks stability during manufacture, shipping, storage, and administration
Solution Approach 1:
The patent applies parameter changes by optimizing the formulation composition including specific buffering agents (Tris, HEPES, phosphate), cryoprotectants (sucrose, trehalose, mannitol), and surfactants (poloxamer 188, polysorbate 80) to achieve enhanced stability. The formulation parameters such as pH (6.5-8.0), ionic strength, and component concentrations are carefully controlled to maintain rAAV potency during extended storage and multiple freeze-thaw cycles.
Solution Approach 2:
The patent employs composite materials by combining multiple excipients and formulation components working synergistically. The composite formulation includes rAAV particles with buffering agents, cryoprotectants, and surfactants that collectively provide protection against degradation during manufacture, shipping, and storage, achieving stability that individual components cannot provide alone.
2Reliability
If the formulation is designed for extended storage at ambient or refrigerated conditions, then stability is improved, but genome release increases
Solution Approach 1:
The patent applies beforehand cushioning by incorporating cryoprotectants (sucrose, trehalose, mannitol) and surfactants into the formulation before storage. These components provide protective cushioning against physical and chemical degradation during freeze-thaw cycles and extended storage, preventing capsid damage and genome release that would otherwise occur under ambient or refrigerated conditions.
3Reliability
If lyophilization is implemented to enhance stability, then storage conditions are improved, but manufacturing complexity increases
Solution Approach 1:
The patent applies preliminary action by optimizing the formulation composition with specific excipients and buffers before the lyophilization process. This preliminary formulation design ensures that the rAAV particles are pre-protected and properly conditioned to withstand the lyophilization stress, enabling successful freeze-drying with minimal additional manufacturing complexity while achieving enhanced storage stability.
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 rAAV potency and stability, with residual moisture content between 1% and 7%, and retains at least 60% relative potency after storage at room temperature for up to 6 months, effectively reducing genome release.
Implementation Method 1
a sugar as a cryoprotective and lyoprotective excipient
Implementation Method 2
a sugar as a cryoprotective and lyoprotective excipient
Implementation Method 3
the formulation is suitable for lyophilization
Implementation Method 4
an amorphous salt comprising sodium citrate
Data Source
Figure 1A~1B
Figure 2
Figure 3
AI summary
Provided herein are formulations comprising recombinant AAV particles. In some embodiments, the formulation is a frozen formulation or a lyophilized formulation. Also provided herein are methods for reducing rAAV genome release from rAAV particles.