Anti-PD-1 Monoclonal Antibody Affinity Maturation via CDR Mutation
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Solution Overview
Problem
Current treatments for various diseases, including cancer and immune system disorders, are limited by the inability to effectively inhibit the PD-1/PD-L1 signaling pathway, which hampers the immune response and tumor cell killing.
Innovation Solution
Development of human anti-PD-1 monoclonal antibodies, such as DFPD1-1, through a method involving biopanning of synthetic ScFv phage libraries, affinity maturation of light and heavy chain CDRs, and cloning into eukaryotic expression vectors, to block the PD-1/PD-L1 interaction and enhance immune response against tumors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional treatments are used, then current therapy options are available, but the PD-1/PD-L1 signaling pathway cannot be effectively inhibited, resulting in limited immune response and tumor cell killing capability
Solution Approach 1:
The patent uses monoclonal antibodies as intermediary substances that specifically bind to PD-1 or PD-L1, blocking their interaction and preventing the inhibitory signaling pathway. This intermediary approach restores T cell function and enhances tumor cell killing without directly attacking the tumor, thereby resolving the contradiction between pathway inhibition and immune response activation.
Solution Approach 2:
The patent replaces conventional chemical therapies or surgery with a biological mechanism-based approach using monoclonal antibodies. The antibodies utilize their binding capability to disrupt the PD-1/PD-L1 interaction, substituting mechanical/chemical treatment with a targeted biological mechanism that restores immune function and enhances tumor killing efficiency.
2Measurement precision
If monoclonal antibodies are developed through synthetic ScFv phage library biopanning, then high affinity anti-PD-1 antibodies can be obtained, but the process requires complex steps including affinity maturation of CDRs and cloning into eukaryotic expression vectors
Solution Approach 1:
The patent segments the antibody development process into distinct stages: initial biopanning of synthetic ScFv phage libraries to obtain candidate antibodies, affinity maturation through CDR mutation to enhance binding affinity, and cloning into eukaryotic expression vectors for production. This segmentation allows systematic optimization at each stage, achieving high affinity while managing process complexity through modular approach.
Solution Approach 2:
The patent employs parameter changes by introducing mutations in the CDR (complementary determining region) segments of the antibody to optimize binding affinity. By systematically altering amino acid sequences in key regions and selecting variants with improved PD-1 binding characteristics, the patent achieves high affinity antibodies through controlled parameter modification rather than random screening.
Data Source
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AI summary
The present invention relates to the technical field of antibody engineering. Provided are a full human anti-PD-1 monoclonal antibody, a preparation method therefor, and uses thereof. Anti-PD-1 monoclonal antibodies are obtained from a fully synthetic antibody library by means of screening, and then light-chain CDR1-3-region and heavy-chain CDR1-3-region mutation construction libraries of the obtained antibody are screened in sequence by means of an affinity maturation technology to obtain a high-affinity anti-PD-1 antibody. The antibody can be used for treating tumors, inflammations, and autoimmune diseases.