Antibody Screening via VH-VL Domain Segmentation
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Solution Overview
Problem
Current methods for selecting antibodies are slow and inefficient in producing antibodies with desired kinetic properties, selectivity, biophysical properties, and immunogenicity necessary for therapeutic use, as they primarily focus on covalent linkage and binding affinity without considering non-covalent interactions between VH and VL domains.
Innovation Solution
The method involves creating a library of unpaired VH or VL domains with diversity in FR1-FR3 regions and selecting pairs based on the strength of non-covalent interactions, in addition to binding affinity, to produce antibodies with improved thermodynamic stability and antigen binding specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If classical methods of immunizing animals are used to obtain antibodies, then antibodies can be obtained with desired antigen binding specificity, but the process is slow and cumbersome
Solution Approach 1:
The patent segments the antibody structure into separate VH and VL domains that can be independently selected and combined. This allows parallel processing of domain libraries rather than working with complete antibody molecules, significantly reducing the time required to generate antibodies with desired specificity while maintaining binding quality.
Solution Approach 2:
The patent performs preliminary selection of VH and VL domains from large libraries before combining them into functional antibodies. By pre-selecting domains with desired properties (specificity, affinity, stability) and then combining them, the overall process time is reduced compared to traditional immunization methods that require waiting for full antibody maturation in vivo.
2Productivity
If in vitro selection techniques such as phage display are used, then the process is faster than animal immunization, but it is still relatively difficult to obtain antibodies with desired kinetic properties, selectivity, biophysical properties, and immunogenicity
Solution Approach 1:
The patent divides the antibody into separate VH and VL domains that can be independently optimized and combined. This segmentation allows for more thorough screening of domain combinations to achieve desired kinetic properties, selectivity, and biophysical characteristics while maintaining fast in vitro selection speeds.
Solution Approach 2:
The patent systematically varies parameters such as domain pairing combinations, CDR region configurations, and framework region selections to optimize multiple antibody properties simultaneously. By changing these parameters across large libraries of VH and VL domains, the method achieves both high productivity and reliable antibody quality with desired kinetic and biophysical properties.
3Ease of manufacture
If VH and VL domains are selected based solely on binding affinity of linked domains to target antigen, then selection is simplified, but the intrinsic thermostability of VH/VL pairs is not optimized
Solution Approach 1:
The patent performs preliminary assessment of VH/VL pair compatibility based on non-covalent interaction strength before final selection based on antigen binding affinity. This two-stage approach maintains simplicity while ensuring that selected pairs have both strong intrinsic stability and desired binding properties.
Solution Approach 2:
The patent introduces an intermediary selection criterion (non-covalent interaction strength between unpaired VH and VL domains) that mediates between the simplicity of affinity-based selection and the need for thermostability optimization. This intermediary step filters domain pairs to ensure stable combinations are selected without significantly complicating the overall process.
Data Source
AI summary
Provided are methods and compositions for the production of novel antibodies that bind specifically to a target antigen. These methods and compositions are particularly useful for producing antibodies having the antigen binding specificity of a reference antibody but with improved properties (e.g., binding affinity, immunogenicity, and thermodynamic stability) relative to the reference antibody.