Rabbit Antibody Humanization via CDR3 Embedding
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Solution Overview
Problem
Current methods for humanizing antibodies, such as CDR grafting and guided selections, are inefficient and limited to processing a few antibodies at a time, requiring extensive trial and error and lacking in high-throughput capabilities for mass humanization of rabbit-derived antibodies.
Innovation Solution
A method for producing nucleic acids encoding immunoglobulin variable domains with rabbit-derived CDR3 amino acid sequences embedded in human framework sequences, allowing for high-throughput and efficient humanization of rabbit antibodies by diversifying CDR1 and CDR2 sequences and using a simplified framework landscape, enabling mass humanization of rabbit antibodies in parallel without determining their amino acid sequences.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If CDR grafting is used to humanize antibodies, then antibody humanization can be achieved, but the process becomes lengthy and requires extensive trial and error
Solution Approach 1:
The patent applies preliminary action by pre-defining the framework regions (FR1-FR4) and CDR1/CDR2 sequences before the humanization process. The method uses predetermined human framework sequences and combines them with rabbit CDR3 sequences in a systematic manner, eliminating the need for extensive trial and error during the actual humanization process.
Solution Approach 2:
The patent segments the antibody into distinct functional regions: framework regions (FR1-FR4) and complementarity determining regions (CDR1-CDR3). By separating the human framework portions from the rabbit CDR3 portions, the method enables modular assembly that reduces the complexity and time required for humanization compared to treating the entire antibody as a unified structure.
2Reliability
If guided selections are used to humanize antibodies, then all traces of non-human origin can be eliminated, but the library size is limited and cannot process infinite input candidates
Solution Approach 1:
The patent creates a universal framework that can accommodate any number of rabbit CDR3 sequences by using the same human framework regions (FR1-FR4) and CDR1/CDR2 construction approach. This standardized template system allows the method to process unlimited input candidates without requiring proportional increases in library size or complexity, as the framework structure remains constant while only the CDR3 variable region changes.
3Reliability
If CDR grafting is used for humanization, then specific antibody humanization can be achieved, but the process is highly individual and depends on structure of the antibody-antigen complex
Solution Approach 1:
The patent applies local quality by maintaining different properties in different regions of the antibody: the framework regions (FR1-FR4) use standardized human sequences with consistent properties, while the CDR3 regions maintain rabbit-derived specificity with variable properties. This localized differentiation allows the method to achieve both standardized processing and specific binding characteristics without requiring complex individual analysis of each antibody-antigen complex.
Data Source
AI summary
The present invention relates to a method for producing a population of 20 or more nucleic acids, each encoding at least one protein comprising at least one immunoglobulin variable domain having a rabbit-derived CDR3 amino acid sequence embedded in essentially human framework sequences, as well as to a population of nucleic acids and a population of proteins relates thereto and uses thereof.