Sortase-Mediated AAV Capsid Conjugation for Targeting
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for conjugating AAV particles with targeting molecules to improve transduction efficiency and specificity are hindered by complex design setups, low modularity, and adverse effects on production yields.
Innovation Solution
Introduction of sortase recognition sequences within specific regions of the AAV capsid proteins, such as the VP1-VP2 transition region, VR-I, VR-IV, and VR-VIII, allows for efficient conjugation of targeting molecules while maintaining structural and functional integrity of the capsid.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If targeting molecules are conjugated to AAV capsids to improve transduction efficiency and specificity, then targeting efficiency is improved, but production yield decreases and manufacturing complexity increases
Solution Approach 1:
The patent introduces sortase recognition sequences into the AAV capsid genome during viral vector production, enabling post-manufacturing conjugation of targeting molecules. This preliminary preparation allows the capsid to be modified after purification without affecting the manufacturing process, thereby maintaining production yield while enabling improved targeting efficiency through subsequent conjugation with targeting ligands.
Solution Approach 2:
The patent uses sortase recognition sequences as intermediary elements that facilitate the conjugation of targeting molecules to AAV capsids. The sortase enzyme recognizes specific sequences in the capsid and enables covalent attachment of targeting ligands, serving as a molecular mediator that connects the capsid to targeting molecules without requiring complex genetic modification or affecting production processes.
2Reliability
If targeting molecules are conjugated to AAV capsids, then transduction specificity is improved, but manufacturing complexity increases
Solution Approach 1:
The patent incorporates sortase recognition sequences into the AAV capsid genome during the initial viral vector production, before purification and formulation. This preliminary genetic modification allows the capsid to retain its native structure and manufacturing properties while enabling subsequent conjugation of targeting molecules through simple enzymatic reactions, thereby improving transduction specificity without increasing manufacturing complexity.
Solution Approach 2:
The patent changes the chemical parameters of the AAV capsid by introducing sortase recognition sequences at specific amino acid positions (such as K258, K321, K490, K507, K527, K532, K544, K549, K556, K620, K640, K649, K665, K692, or K706 in AAV2). These parameter changes enable covalent attachment of targeting ligands through sortase-mediated conjugation, improving transduction specificity while maintaining compatibility with existing manufacturing processes.
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
This approach enables the production of AAV particles with enhanced targeting specificity and efficiency, while maintaining similar production efficiencies compared to parental AAV serotypes, thus overcoming the limitations of existing methods.
Implementation Method 1
sortase recognition sequences can be introduced internally in capsid sequences of AAV particles to enable conjugation of targeting molecules with the AAV capsids
Data Source
AI summary
The invention relates to an adeno-assisted virus (AAV) VP1, VP2 or VP3 capsid protein characterized in that in one or more of the VP1-VP2 transition region, VR-I region, VR-IV region and VR-VIII region a sortase recognition is inserted, wherein n and m range from 0 to 25, and wherein X is any natural amino acid.