AAV Capsid Engineering for High-Titer Production

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Solution Overview

Problem

Current methods for producing adeno-associated viruses (AAVs) face challenges in achieving high titers, which limits their efficiency and application in gene transfer and therapeutic uses.

Innovation Solution

Designing AAVs with a capsid polypeptide containing a specific amino acid sequence (DPIVMIDNDKPIT or variants) that allows for covalent bonding with a conjugating polypeptide, enabling the formation of high-titer preparations of AAVs by linking two or more AAVs covalently, thereby enhancing their production and utilization.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If current AAV production methods are used, then AAV preparations can be produced, but the titer of functional AAVs is limited and cannot achieve high titers greater than 10^14

Engineering Contradiction:

Improvetiter of functional AAVsVSAvoidproduction efficiency

Core Design Contradiction:

Quantity of substanceVSProductivity

Solution Approach 1:

The patent modifies the capsid polypeptide by introducing a specific amino acid segment (DPIVMIDNDKPIT or variants) at defined positions, changing the chemical parameters of the AAV structure to enable covalent bonding capability, which directly enables high-titer production

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent creates composite AAV structures by covalently linking multiple AAVs together through the modified capsid polypeptides, forming multi-AAV complexes that achieve high titers greater than 10^14 functional AAVs per preparation

Inventive Principle:

Principle #40Composite materials

2Quantity of substance

If AAVs are designed with specific amino acid segments for covalent bonding, then high-titer preparations can be produced, but the capsid structure becomes more complex

Engineering Contradiction:

Improvetiter of functional AAVsVSAvoidcapsid polypeptide structure

Core Design Contradiction:

Quantity of substanceVSDevice complexity

Solution Approach 1:

The patent introduces the bonding-enabled amino acid segment at specific local positions (e.g., positions 453 or 588 of reference VP1 polypeptide) rather than throughout the entire capsid structure, maintaining local functionality while enabling global high-titer production

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The patent segments the capsid polypeptide by inserting a distinct amino acid segment (DPIVMIDNDKPIT) that serves as a functional module for covalent bonding, separating the bonding function from the rest of the capsid structure

Inventive Principle:

Principle #1Segmentation

Data Source

PatentUS20220348613A1Adeno-associated viruses and methods and materials for making and using adeno-associated viruses

Publication Date: 2022.11.03 RGT UNIV OF CALIFORNIA

AI summary

This document provides AAVs and methods and materials for making and using AAVs. For example, AAVs containing a capsid polypeptide that includes an amino acid segment having a DPIVMIDNDKPIT sequence (or a variant thereof) are provided. This document also provides compositions containing an AAV described herein, nucleic acid molecules encoding an AAV described herein, conjugating polypeptides, nucleic acid molecules encoding a conjugating polypeptide described herein, and methods for making a composition that includes two or more different AAVs covalently linked together.