Variable Domain Aggregation Resistance via CDR2 Charge

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Solution Overview

Problem

Current methods for creating aggregation-resistant antibody variable domains are limited by their applicability across various proteins, often requiring detailed structural analysis and specialized knowledge, and can result in reduced antigen affinity or immunogenicity.

Innovation Solution

Introduction of negatively charged amino acids into the complementarity determining region 2 (CDR2) of the variable domain, specifically at positions between residues 49 and 56, to enhance aggregation resistance without compromising antigen binding affinity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If variable domains are stabilized to resist aggregation, then aggregation resistance is improved, but antigen binding affinity may be reduced

Engineering Contradiction:

Improveaggregation resistanceVSAvoidantigen binding affinity

Core Design Contradiction:

ReliabilityVSManufacturing precision

Solution Approach 1:

The patent introduces negatively charged amino acids (aspartic acid or glutamic acid) at specific positions (49-56) in the CDR2 region of the variable domain. This chemical parameter change increases aggregation resistance through electrostatic repulsion while the patent specifically selects positions that do not interfere with antigen binding, thus maintaining binding affinity.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The modification is localized to specific positions within the CDR2 region (positions 49-56) rather than throughout the entire variable domain. This local introduction of negative charges provides aggregation resistance at the surface while preserving the structural integrity and antigen-binding capability of the core variable domain structure.

Inventive Principle:

Principle #3Local quality

2Reliability

If detailed structural analysis is performed to create aggregation-resistant variable domains, then aggregation resistance is improved, but method complexity increases

Engineering Contradiction:

Improveaggregation resistanceVSAvoidmethod complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent establishes specific parameter guidelines: introducing negatively charged amino acids at positions 49-56 in CDR2. This provides a clear, actionable protocol that reduces method complexity compared to performing detailed structural analysis of each variable domain, while still achieving aggregation resistance through the defined parameter changes.

Inventive Principle:

Principle #35Parameter changes

3Reliability

If negatively charged amino acids are introduced into CDR2, then aggregation resistance is improved, but immunogenicity may increase

Engineering Contradiction:

Improveaggregation resistanceVSAvoidimmunogenicity

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The negative charges are introduced locally at positions 49-56 in CDR2, which are typically surface-exposed regions. This localized modification provides aggregation resistance while the patent selects positions that are less likely to be recognized as immunogenic epitopes, balancing aggregation resistance with reduced immunogenicity.

Inventive Principle:

Principle #3Local quality

Data Source

PatentUS11332522B2Modified variable domain molecules and methods for producing them

Publication Date: 2022.05.17 GARVAN INSTITUTE OF MEDICAL RESEARCH

AI summary

The present disclosure provides an isolated, engineered or non-naturally occurring protein comprising an antibody light chain variable domain (VL) which may comprise at least one negatively charged amino acid positioned between residues 49 to 56 according to the numbering system of Kabat, the protein capable of binding specifically to an antigen.