αVβ3 Integrin Single-Domain Antibody for Tissue-Permeable Detection
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Solution Overview
Problem
Existing antibodies are too large and chemically unstable to be useful in living cells, limiting their ability to observe protein folding and interactions in real time, and there is a need for effective tools to detect and diagnose angiogenesis-related diseases.
Innovation Solution
Development of an αvβ3 integrin targeting single-domain antibody, encoded by specific base sequences, which is smaller, more stable, and can be easily expressed in bacteria and yeast, allowing for detection and diagnosis of angiogenesis through recombinant vectors and microorganisms.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional antibodies are used for detecting protein interactions in living cells, then they can bind to target antigens with high specificity, but their large size (150 kDa) and chemical instability prevent them from being useful in living cells and migrating into cells
Solution Approach 1:
The patent extracts only the essential binding domain (variable heavy chain VHH) from the complete antibody structure, creating a single-domain antibody that retains antigen-binding capability while removing the bulk of the molecular weight. This extracted VHH domain (12-13 kDa) can penetrate cell membranes and function within living cells, resolving the contradiction between binding reliability and molecular size.
Solution Approach 2:
The patent segments the conventional antibody structure into separate functional domains, isolating the variable heavy chain (VHH) as an independent functional unit. This segmentation allows the VHH to operate autonomously with cell-permeable properties while maintaining the specific binding function, thus solving the size-stability problem of conventional antibodies.
2Measurement precision
If conventional antibodies are used for observing protein folding and interactions in real time, then they can provide detailed binding information, but their large size and instability make real-time observation in living cells impossible
Solution Approach 1:
The patent employs single-domain antibodies that are chemically stable yet small enough to function transiently within living cells for real-time observation. These VHH antibodies can be expressed in bacteria and yeast, allowing rapid production and deployment for time-sensitive measurements of protein folding and interactions without requiring long-term stability.
Solution Approach 2:
The patent changes the physical-chemical parameters of the antibody by reducing its molecular weight from 150 kDa to 12-13 kDa while maintaining binding affinity. This parameter change enables the antibody to diffuse into cells and remain stable under intracellular conditions, facilitating real-time measurement of dynamic protein processes.
3Weight of moving object
If recombinant antibodies are used instead of conventional antibodies, then their size is reduced (25-50 kDa), but they are still too large to easily migrate into cells compared to single-domain antibodies
Solution Approach 1:
The patent extracts only the essential VHH domain from the recombinant antibody structure, creating an even smaller single-domain antibody (12-13 kDa) that can easily migrate into cells. This extraction removes unnecessary structural elements while preserving the binding function, achieving superior cell permeability compared to full recombinant antibodies.
4Ease of operation
If single-domain antibodies are used for detecting angiogenesis, then they can penetrate tissues and bind to αvβ3 integrin, but there is a need for effective tools to detect and diagnose angiogenesis-related diseases
Solution Approach 1:
The patent develops single-domain antibodies with universal applicability for detecting various angiogenesis-related targets including αvβ3 integrin. These VHH antibodies can be used in multiple diagnostic formats and applications, providing a versatile tool for detecting and diagnosing angiogenesis-related diseases through their excellent tissue permeability and binding specificity.
Data Source
AI summary
Disclosed are an αvβ3 integrin targeting single-domain antibody and various applications thereof. The αvβ3 integrin targeting single-domain antibody exhibits high binding ability to αvβ3 integrin related to angiogenesis, excellent tissue permeability, and biostability compared to conventional antibodies. Further, the single-domain antibody may be combined with fluorescent particles and thus may be easily measured in vitro, in vivo or ex vivo, and may be effective in detecting angiogenesis and diagnosing angiogenesis related diseases, therefore it may be usefully used in related industries.