Multispecific Antibody CH1-CL Interface Engineering
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Solution Overview
Problem
The production and development of bispecific antibodies are hindered by challenges such as light chain mispairing, which affects the stability and pharmacokinetic properties of these molecules, and existing strategies are technically difficult to implement and lack universality.
Innovation Solution
The introduction of non-natural disulfide bonds and electrostatically complementary amino acids at specific positions in the CH1/CL interface of multispecific antibodies to enhance correct pairing and stability, including substitutions like C220A, F170C, and T139R, which promote electrostatic interactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If existing strategies (common light chains, two-in-one antibodies, IgG/TCR, Crossmab, DuetMab, computer-aided design) are used to avoid light chain mispairing, then mispairing is reduced, but the structural adjustments alter stability, immunogenicity or pharmacokinetic properties
Solution Approach 1:
The patent applies parameter changes by systematically modifying amino acid positions at the CH1/CL interface (positions 128, 129, 131, 141, 170, 171 in CH1 and corresponding positions in CL) to create orthogonal interactions. These parameter changes enable correct heavy chain-light chain pairing while preserving molecular stability, immunogenicity, and pharmacokinetic properties through optimized interface interactions rather than structural alterations.
2Reliability
If existing strategies are used to avoid light chain mispairing, then mispairing is reduced, but the implementation is technically difficult and lacks universality
Solution Approach 1:
The patent achieves universality by developing a standardized orthogonal interface system that can be applied across different antibody formats and production methods. The defined amino acid substitutions at specific CH1/CL interface positions create a universal solution for preventing light chain mispairing that works regardless of the specific production strategy employed, making the approach broadly applicable in the field.
3Manufacturing precision
If natural disulfide bonds are removed and non-natural disulfide bonds are introduced at CH1/CL interface, then correct pairing is enhanced, but structural complexity increases
Solution Approach 1:
The patent applies local quality by introducing specific amino acid substitutions only at defined positions within the CH1/CL interface region (positions 128, 129, 131, 141, 170, 171 in CH1 and corresponding positions in CL). This localized modification approach enhances pairing accuracy through orthogonal interactions at the interface while leaving the rest of the antibody structure unchanged, thereby minimizing overall structural complexity.
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 increases the proportion of correct heavy chain-light chain pairing, improving the yield and stability of multispecific antibodies, thereby addressing the mispairing issues and enhancing their biological functions.
Implementation Method 1
introducing a non-natural disulfide bond into the interface
Implementation Method 2
introducing a pair of electrostatically complementary amino acids into the CH1/CL interface
Data Source
AI summary
Provided is a multispecific antigen binding protein. Provided is a multispecific antigen binding protein comprising one or more amino acid substitutions at CH1 and CL, a composition comprising same, a preparation method therefor, and a medical use thereof. The specific antigen binding protein effectively reduces the mismatching of light chains.