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A phage display approach for rapid antibody humanization: designed combinatorial V gene libraries
C Rader1, D A Cheresh, C F Barbas
1Skaggs Institute for Chemical Biology and Department of Molecular Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Summary
A novel antibody humanization strategy retains critical mouse antibody sequences within complementarity-determining regions (CDRs) while replacing others with human sequences. This method rapidly generates high-affinity, specific humanized antibodies for potential cancer therapy.
Area of Science:
- Immunology
- Biotechnology
- Molecular Biology
Background:
- Antibody humanization is crucial for reducing immunogenicity in therapeutic antibodies.
- Existing methods may not optimally preserve critical antibody-binding properties.
Purpose of the Study:
- To develop and validate a new phage display-based strategy for antibody humanization.
- To humanize the anti-integrin antibody LM609 for potential anti-cancer applications.
Main Methods:
- Incorporation of parental rodent antibody's key recognition sequences (CDRs) into a phage display system.
- Selection of human sequences from phage-displayed libraries to replace non-CDR regions.
- Application to mouse monoclonal antibody LM609 targeting human integrin alphav beta3.
Main Results:
- Rapid antibody humanization with minimal retention of original mouse sequences (limited to CDRs).
- Generation of multiple humanized antibody variants simultaneously.
- Achieved high or improved antibody affinities compared to the original.
- Maintained original antigen and epitope specificity.
- Demonstrated potential for large-scale expression and therapeutic use, including avoiding anti-idiotypic responses.
Conclusions:
- The described strategy offers an efficient and effective method for antibody humanization.
- It yields humanized antibodies with preserved or enhanced functionality and specificity.
- This approach has significant implications for developing therapeutic antibodies, particularly for long-term cancer treatment.