AAV Capsid Composition for Reduced Deamidation During Storage
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Solution Overview
Problem
There is a need for AAV-based constructs that maintain stable receptor binding, avoid neutralizing antibodies, and retain purity during storage, as current AAV gene therapy vectors lack understanding of post-translational modifications (PTMs) that impact function and stability.
Innovation Solution
A composition of recombinant adeno-associated virus (rAAV) with heterogeneous populations of VP1, VP2, and VP3 proteins containing specific amino acid modifications, such as deamidated asparagines and glutamines, to stabilize the capsid and reduce deamidation, along with a vector genome for gene expression.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If AAV gene therapy vectors are manufactured and stored, then the vectors can be used for gene delivery, but the capsid proteins undergo deamidation which reduces stability and purity
Solution Approach 1:
The patent applies parameter changes by modifying amino acid residues in the capsid protein sequence, specifically targeting asparagine and glutamine residues that are prone to deamidation. By changing these residues to alternative amino acids (e.g., substituting asparagine with aspartic acid or glutamine with glutamic acid), the patent alters the chemical parameters of the capsid protein to resist deamidation during storage, thereby maintaining compositional stability while enabling long-term storage of AAV vectors
2Reliability
If AAV vectors are produced with modified capsid proteins to reduce deamidation, then stability and purity are improved, but the complexity of vector production increases
Solution Approach 1:
The patent applies preliminary action by incorporating the desired amino acid modifications directly into the capsid protein gene sequence during the vector design phase. This preliminary genetic engineering ensures that the modified capsid proteins are produced with reduced deamidation propensity from the outset, eliminating the need for post-production modification steps and simplifying the overall production process while maintaining high vector stability and reliability
3Productivity
If AAV vectors maintain high purity during storage, then transduction efficiency is improved, but the cost of manufacturing and quality control increases
Solution Approach 1:
The patent applies self-service by engineering the capsid proteins to inherently resist deamidation through amino acid substitution. This self-protective modification allows the AAV vectors to maintain high purity during storage without requiring complex external stabilization measures, expensive protective formulations, or frequent quality intervention, thereby preserving transduction efficiency while reducing manufacturing and quality control costs
Data Source
AI summary
A recombinant adeno-associated virus (rAAV) vector comprising an AAV capsid having a heterogeneous population of vp1 proteins, a heterogeneous population of vp2 protein and a heterogeneous population of vp3 proteins. The capsid contains modified amino acids as compared to the encoded VP1 amino acid sequence, the capsid containing highly deamidated asparagine residues at asparagine-glycine pair, and further comprising multiple other, less deamidated asparagine and optionally glutamine residues.