Bispecific Antibody Pairing Using Kappa-Lambda Fab Engineering

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Solution Overview

Problem

Efficient production of bispecific antibodies that bind to two different epitopes is challenging due to the promiscuous pairing of antibody heavy and light chains, particularly when one parent antibody has a kappa light chain and the other has a lambda light chain, which complicates homogeneous pairing and manufacturability.

Innovation Solution

Development of multispecific antigen-binding polypeptides with engineered immunoglobulin lambda and kappa light chains, incorporating specific amino acid modifications to promote preferential pairing of heavy and light chains, ensuring stable and efficient formation of Fab regions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If conventional bispecific antibody production methods are used with kappa and lambda light chains, then two different epitopes can be targeted, but heavy chain-light chain pairing becomes scrambled and manufacturing becomes difficult

Engineering Contradiction:

Improveability to bind two different epitopesVSAvoiddifficulty of producing homogeneous bispecific antibodies

Core Design Contradiction:

Adaptability or versatilityVSEase of manufacture

Solution Approach 1:

The patent introduces specific amino acid modifications at the heavy chain-light chain interface to create asymmetric pairing properties. By modifying residues at positions 100-150 of the heavy chain and corresponding positions on the light chain, the invention creates localized changes that enable H1 to preferentially bind L1 and H2 to preferentially bind L2, thereby resolving the scrambling issue while maintaining the ability to target two different epitopes

Inventive Principle:

Principle #3Local quality

Solution Approach 2:

The invention employs asymmetric amino acid substitutions at the heavy chain-light chain interface to create directional pairing specificity. The modifications create an asymmetric binding landscape where each heavy chain has a unique preferred light chain partner, preventing promiscuous pairing and enabling homogeneous production of bispecific antibodies with two different light chain types

Inventive Principle:

Principle #4Asymmetry

2Manufacturing precision

If amino acid modifications are introduced to promote preferential pairing, then manufacturing precision improves, but the complexity of the antibody construct increases

Engineering Contradiction:

Improvepairing specificity of heavy and light chainsVSAvoidnumber of amino acid modifications required

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent modifies specific parameters of the heavy chain and light chain sequences by introducing amino acid substitutions at defined positions (100-150 of heavy chain and corresponding light chain positions). These parameter changes alter the binding interface properties to create preferential pairing without requiring extensive modifications throughout the entire antibody structure, thus achieving high pairing specificity with controlled complexity

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS12540198B2Antigen-binding polypeptide constructs comprising kappa and lambda light chains and uses thereof

Publication Date: 2026.02.03 ZYMEWORKS BC INC

AI summary

Provided herein are multispecific antigen-binding polypeptide constructs comprising at least two different heterodimers, each comprising a heavy chain and a light chain. At least one heterodimer comprises a Fab region comprising a lambda light chain and at least one heterodimer comprises a Fab region comprising a kappa light chain. One or more of the immunoglobulin heavy and light chains that form the antigen-binding polypeptide construct comprise amino acid modifications that promote correct pairing between the heavy and light chains to form the desired multispecific antigen-binding polypeptide construct. The amino acid modifications may be in the CH1 and/or CL domains, in the VH and/or VL domains, or a combination thereof.