Human Naïve Antibody Library via Phage Yeast Display
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Solution Overview
Problem
Current methods for generating human antibody libraries face limitations such as immunogenicity, pharmacokinetic and safety issues with chimeric and non-human antibodies, and challenges in generating antibodies against self-antigens due to toxicity and non-immunogenicity in murine hybridoma technology, as well as limited control over epitope formation and antigen specificity.
Innovation Solution
A method for generating a highly diverse human naïve antibody library using a combination of phage and yeast antibody surface display technologies, involving processing biological samples to isolate and amplify nucleic acids, cloning into phage or yeast vectors, and screening against antigens to select antibodies with desired functional properties, employing degenerate primers and multiple rounds of selection to achieve high affinity and specificity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If chimeric or non-human antibodies are used, then antibody production is achieved, but immunogenicity and safety issues arise
Solution Approach 1:
The patent changes the species origin parameter of the antibody from non-human or chimeric to fully human by using human B-cell lymphoma cell lines (such as CHO-K1, CHO-DG44, CHO-ZB1) as expression systems. This parameter change eliminates immunogenicity while maintaining antibody production efficacy, as human antibodies do not trigger immune responses in human patients.
2Reliability
If murine hybridoma technology is used, then antibody generation is possible, but control over epitope formation and antigen specificity is limited
Solution Approach 1:
The patent segments the antibody generation process into distinct modular steps: (1) selection of human B-cell lymphoma cell lines expressing desired antibody specificities, (2) isolation and characterization of antibody genes, (3) cloning into appropriate vectors, and (4) expression in human cell lines. This segmentation allows precise control over each step, particularly enabling control over epitope formation by selecting cell lines with known antigen specificities before proceeding to cloning and expression.
3Adaptability or versatility
If traditional immunization methods are used, then antibody diversity is generated, but safety and pharmacokinetic properties are compromised
Solution Approach 1:
Instead of using traditional immunization methods that require animal models and carry safety risks, the patent copies the natural human antibody repertoire directly from human B-cell lymphoma cell lines. These cell lines naturally express diverse human antibodies, allowing the patent to obtain human antibody diversity without animal immunization, thereby ensuring safety and optimal pharmacokinetic properties from the outset.
4Reliability
If antibodies are generated against toxic antigens using murine hybridoma, then antibody production is achieved, but antigen toxicity and non-immunogenicity in mice limit utility
Solution Approach 1:
The patent uses human B-cell lymphoma cell lines as an intermediary system that can safely handle and express antibodies against toxic or self-antigens. These cell lines serve as a bridge between the toxic antigen and the final therapeutic antibody product, allowing antibody generation against targets that would be toxic or non-immunogenic in murine hybridoma systems, thereby expanding applicability to toxic self-antigens and human-specific targets.
Data Source
AI summary
The present disclosure relates to a method of generating an antibody library, not limiting to a human naïve antibody gene expression library encompassing a pool of nucleic acid sequences derived from a natural antibody repertoire comprising humoral immunity from healthy and genetically diverse human populations. More specifically, the method employs a combination of phage and/or yeast antibody surface display concept which allows to screen large antibody library size and facilitates better folding of antibody structure. The present disclosure also relates to a human naïve antibody library generated by employing the process of the present disclosure, a set of primers employed in the method and application(s) of said antibody library.