Humanized CD3 Antibody Framework Optimization
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Solution Overview
Problem
Current anti-CD3 antibodies, such as murine antibodies, often induce a human anti-mouse antibody (HAMA) response, leading to adverse reactions in humans, and humanized antibodies face challenges like decreased affinity, stability issues, and low yield, making them ineffective for therapeutic use.
Innovation Solution
Development of humanized antibodies with specific framework regions and complementarity-determining regions (CDRs) sequences that target CD3 in primates, including humans and monkeys, with optimized heavy and light chain variable regions to enhance affinity, stability, and activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If murine anti-CD3 antibodies are used, then antibody activity against CD3 is achieved, but human anti-mouse antibody (HAMA) response is induced causing adverse reactions
Solution Approach 1:
The patent applies parameter changes by humanizing the antibody sequence - specifically changing the species origin parameter from murine to human through sequence optimization. The humanized antibody maintains CD3 binding activity while eliminating the foreign antigenic determinants that trigger HAMA response, thus resolving the contradiction between maintaining antibody activity and reducing adverse immune reactions
Solution Approach 2:
The patent creates a humanized version of the murine antibody by copying and adapting the antigen-binding specificities while using human framework regions. This allows the antibody to function as if it were naturally human-derived, avoiding the immune system's recognition of foreign mouse protein structures while preserving the therapeutic anti-CD3 activity
2Object-affected harmful factors
If murine antibodies are humanized to reduce HAMA response, then adverse reactions are reduced, but antibody affinity and stability decrease
Solution Approach 1:
The patent applies local quality by selectively humanizing only the framework regions while preserving the critical complementarity-determining regions (CDRs) that confer high affinity binding. This localized approach allows different parts of the antibody to have different degrees of humanization - the framework is humanized to reduce immunogenicity, while the CDRs retain their original sequences to maintain binding affinity and stability
Solution Approach 2:
The patent segments the antibody into functional domains with different humanization strategies. The framework regions are humanized to reduce HAMA response, while the CDR regions are preserved to maintain affinity. This segmentation allows independent optimization of immunogenicity and binding properties without compromising either aspect
3Object-affected harmful factors
If humanized antibodies are developed to avoid HAMA response, then safety is improved, but manufacturing yield is low
Solution Approach 1:
The patent optimizes the humanized antibody sequence parameters to improve expression characteristics. By carefully selecting framework region sequences and optimizing codon usage, the patent enhances the manufacturability and yield of the humanized antibody while maintaining the safety benefits of reduced immunogenicity
Data Source
AI summary
Provided in the present application is a humanized and multifunctional CD3 antibody; a heavy chain variable region of the CD3 antibody comprised herein contains the amino acid sequence of any one of SEQ ID NOs: 1-3 or a variant sequence thereof; and a light chain variable region thereof contains the amino acid sequence of any one of SEQ ID NOs: 26-28 or a variant sequence thereof. Further provided in the present application is a multifunctional antibody comprising (a) a light chain-heavy chain pair that has specificity for tumor cells or microorganisms; and (b) a fusion peptide that comprises a single-chain variable fragment and an Fc fragment having a CH2 domain and/or a CH3 domain, the fusion peptide having specificity for immune cells. The antibodies provided in the present application have improved biological activity, thermal stability and/or acid resistance.