Antibody Library Construction via SynNotch Segmentation
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Solution Overview
Problem
Current antibody screening technologies are limited in their ability to efficiently screen polyclonal antibodies against complex antigens, particularly in the context of tumor therapy, where tumor antigens are complex, diverse, and mutable, and the available targets are limited.
Innovation Solution
The development of an antibody library construction method based on the synNotch system, which controls gene expression in cells. This method involves modifying the extracellular recognition domain of the synNotch system into an extracellular antibody library coding domain and changing the regulated target gene into a screening marker gene, thereby creating an antigen-activated antibody screening system capable of screening polyclonal antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If conventional antibody screening technologies are used, then the screening process is simple, but the ability to screen polyclonal antibodies against complex antigens is limited
Solution Approach 1:
The patent segments the antibody library construction into distinct functional modules: synNotch system components (extracellular recognition domain, Notch core domain, intracellular transcription domain) and screening components (CIS activator, screening marker gene). This modular segmentation enables independent optimization of each component and facilitates the screening of diverse polyclonal antibodies against complex tumor antigens while maintaining manageable system complexity
Solution Approach 2:
The patent introduces the synNotch system as an intermediary mechanism between the antibody library and the screening process. The synNotch system acts as a mediator that controls gene expression in response to antigen binding, enabling indirect screening of polyclonal antibodies through the expression of screening marker genes rather than direct detection, thus improving adaptability while managing complexity
2Productivity
If high throughput screening is implemented, then the screening efficiency is improved, but the complexity of the screening system increases
Solution Approach 1:
The patent implements self-service mechanisms where the synNotch system automatically activates the screening marker gene expression in response to antigen binding, eliminating the need for complex external detection systems. The system performs the screening function through its own internal mechanisms, improving throughput efficiency while minimizing the addition of external complexity
Solution Approach 2:
The patent utilizes parameter changes in the synNotch system activation state to control the expression of screening marker genes. By changing the activation parameters (antigen presence, signal transduction), the system achieves high throughput screening through controlled gene expression rather than complex detection procedures, thereby improving productivity without proportionally increasing system complexity
Data Source
AI summary
Disclosed by the present application are an antibody library construction method and an application thereof. The method comprises the following steps: inserting a first element and a second element into a same vector or different vectors, and transfecting the vectors into the cells to obtain an antibody expression cell library, i.e., the antibody library. The first element comprises CIS activators and selection marker genes; the second element comprises extracellular antibody library coding domain, Notch nuclear structure domain and intracellular transcription structure domain.