Modified IgG4 Antibody Hinge Substitutions for Chromatography

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Therapeutic IgG4 antibodies with serine-to-proline substitutions in the core-hinge region exhibit undesirable bioanalytical and bioprocessing behaviors, including two-peak elution behavior on cation exchange chromatography, which complicates bioanalytical method development and bioprocessing, and can lead to unpredictable Fab-arm exchange and bispecific antibody formation.

Innovation Solution

Designing modified IgG4 antibodies with specific substitutions in the CH1 or hinge region, such as lysine to proline at position 196, serine to proline at position 217, glycine to threonine at position 220, proline to histidine at position 224, or proline to threonine at position 225, to improve bioanalytical and bioprocessing properties, including single-peak elution behavior on cation exchange chromatography.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If serine-to-proline substitution is made at position 228 in the hinge region to prevent Fab-arm exchange, then therapeutic activity is improved, but bioanalytical and bioprocessing properties deteriorate due to two-peak elution behavior on cation exchange chromatography

Engineering Contradiction:

Improvetherapeutic activityVSAvoidbioprocessing properties

Core Design Contradiction:

ReliabilityVSEase of manufacture

Solution Approach 1:

The patent changes the amino acid substitution parameter from S228P to alternative substitutions (K196P, S217P, G220T, P224H, or P225T) in the CH1 or hinge region. These parameter changes maintain the therapeutic function of preventing Fab-arm exchange while eliminating the two-peak elution behavior on cation exchange chromatography, thereby improving bioprocessing properties.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent applies local quality by making site-specific amino acid substitutions at different positions (K196, S217, G220, P224, or P225) rather than a uniform approach. Each substitution location has been optimized to achieve the dual benefit of preventing Fab-arm exchange while maintaining single-peak elution behavior, allowing localized modification to resolve the contradiction.

Inventive Principle:

Principle #3Local quality

2Reliability

If serine-to-proline substitution is made at position 228 in the hinge region to prevent Fab-arm exchange, then therapeutic activity is improved, but bioanalytical method development becomes complicated due to two-peak elution behavior

Engineering Contradiction:

Improvetherapeutic activityVSAvoidbioanalytical method complexity

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent modifies the amino acid substitution parameter from S228P to alternative substitutions (K196P, S217P, G220T, P224H, or P225T). These parameter changes eliminate the two-peak elution behavior on cation exchange chromatography, simplifying bioanalytical method development while preserving the therapeutic function of preventing Fab-arm exchange.

Inventive Principle:

Principle #35Parameter changes

3Reliability

If serine-to-proline substitution is made at position 228 in the hinge region to prevent Fab-arm exchange, then therapeutic activity is improved, but unpredictable bisspecific antibody formation occurs

Engineering Contradiction:

Improvetherapeutic activityVSAvoidantibody composition stability

Core Design Contradiction:

ReliabilityVSStability of the object's composition

Solution Approach 1:

The patent changes the amino acid substitution parameter from S228P to alternative substitutions (K196P, S217P, G220T, P224H, or P225T). These parameter changes provide more stable and predictable antibody composition by eliminating unpredictable Fab-arm exchange and bispecific antibody formation, while maintaining the therapeutic function.

Inventive Principle:

Principle #35Parameter changes

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 modified IgG4 antibodies demonstrate improved bioanalytical and bioprocessing properties, such as single-peak elution on CEX resin, preventing Fab-arm exchange, and maintaining binding activity, thus facilitating robust manufacturing and bioanalytical method development.

Implementation Method 1

the IgG4 antibody binds to a target molecule selected from CD137, CXCR4, eTau, CSF1R, Lag3, PD1, PDL1 or KIR

Methodology Applied

Scientific EffectAntigen-antibody binding:

Implementation Method 2

the improved bioanalytical properties comprise a single-peak elution behavior on analytical cation exchange chromatography (CEX) rein

Methodology Applied

Scientific EffectCation exchange chromatography: Ion Exchange

Data Source

PatentUS11542337B2Therapeutic immunoglobulin G4 for improved bioanalytical and bioprocessing properties

Publication Date: 2023.01.03 BRISTOL MYERS SQUIBB CO

AI summary

In certain embodiments, the disclosure provides an IgG4 antibody comprising a heavy chain and a light chain, wherein the heavy chain comprises: (a) a modified IgG4 CH1 region having a substitution of the lysine residue at position 196; or (b) a modified IgG4 hinge region having a substitution of the serine residue at position 217, the glycine residue at position 220, the proline residue at position 224 or the proline residue at position 225. Preferably, the IgG4 antibody further comprises a substitution of the serine residue at position 228 in the heavy chain hinge region.