Virion Recombination for Parallel Binding Moiety Identification
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Solution Overview
Problem
Current methods face challenges in identifying multiple binding moieties for multiple antigens in a single reaction, making it difficult to efficiently identify cognate pairs.
Innovation Solution
The method involves using a population of cells with surface antigens and attachment-defective virions displaying transgenic viral surface binding moieties, allowing selective infection and recombination between nucleic acids with specific motifs to generate recombinant products that identify cognate binding moieties.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional methods (phage display, ribosome display, etc.) are used to identify binding moieties, then individual binding moieties can be identified, but multiple binding moieties of multiple antigens cannot be identified in a single reaction
Solution Approach 1:
The patent combines multiple identification reactions into a single multi-well plate system where different antigens are presented in different wells, allowing simultaneous identification of multiple binding moiety-antigen pairs in parallel, thereby increasing throughput without proportionally increasing complexity
Solution Approach 2:
The invention creates a universal identification platform that can handle multiple antigens and binding moieties using the same basic methodology and reagents, allowing the system to perform multiple identification functions simultaneously through standardized procedures
2Reliability
If site-specific recombination motifs are integrated into virion genomes, then recombination efficiency between virion and cell nucleic acids is improved, but the complexity of viral vector design increases
Solution Approach 1:
The recombination motifs are pre-integrated into the virion genome during viral vector construction, so that when the virion infects the cell, recombination can occur immediately without requiring additional preparation steps, thereby ensuring efficient and reliable recombination
Solution Approach 2:
The site-specific recombination motifs act as intermediary sequences that facilitate the joining of virion nucleic acid and cell nucleic acid through specific recombination enzymes, making the recombination process more efficient and reliable
3Adaptability or versatility
If multiple transgenic viral surface binding moieties are displayed on virions, then the ability to identify multiple cognate pairs is improved, but the difficulty of managing and tracking individual binding moieties increases
Solution Approach 1:
The patent uses nucleic acid sequences that encode the viral surface binding moieties as molecular copies or identifiers. By sequencing these nucleic acids after recombination, the invention enables easy tracking and identification of which binding moieties are associated with which antigens, solving the tracking difficulty
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enables the simultaneous identification of multiple binding moieties for multiple antigens, increasing transformation efficiency and recombination rates, and facilitating the detection of cognate pairs through functional marker expression.
Implementation Method 1
incubating the population of cells under conditions sufficient to allow recombination between the first and second recombination motifs in infected cells, thereby generating a recombinant product
Implementation Method 2
binding of the cell surface antigen and the cognate binding moiety results in selective infection by the bound virion of the cell including the cell surface antigen of the cognate binding moiety
Data Source
AI summary
The identification of binding moieties capable of selectively interacting with one or more target antigens is of scientific, medical, and commercial value. Disclosed herein are methods and compositions for the identification, labeling and/or retrieval of cognate binding moieties.