Polyclonal Antibody Mixtures for Epitopic Coverage Normalization
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Solution Overview
Problem
Current antibody display technologies face challenges in comprehensively interrogating the full repertoire of epitopes on antigens, particularly when there is little prior knowledge of epitopic diversity, leading to time-consuming and laborious processes dominated by binders to dominant epitopes, making it difficult to isolate antibodies targeting rare epitopes.
Innovation Solution
The development of methods and reagents that broaden, normalize, and reduce bias in epitopic coverage by using polyclonal mixtures of antibodies, which involve contacting antigens with libraries of antibodies, collecting binders, and re-interrogating with polyclonal mixtures to enrich for rare or under-represented epitopes without prior knowledge of epitopic coverage or diversity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional antibody display technologies are used to screen for antibodies against an antigen, then the process can identify antibodies binding to dominant epitopes, but it becomes time-consuming and laborious to isolate antibodies targeting rare epitopes due to selection bias towards dominant epitopes
Solution Approach 1:
The method performs preliminary actions by first identifying dominant epitopes and their corresponding antibodies, then using these to block dominant epitopes in subsequent screening rounds. This preliminary blocking action prevents selection bias before the actual rare epitope antibody isolation begins, thereby reducing the time needed to isolate rare epitope antibodies while maintaining accurate epitopic coverage.
Solution Approach 2:
The patent uses polyclonal antibody mixtures as intermediaries to bridge the gap between dominant and rare epitope detection. These mixtures contain antibodies against multiple epitopes and are used to block dominant epitopes during screening, allowing rare epitope antibodies to be identified without the time-consuming process of direct screening against all epitopes simultaneously.
2Productivity
If antibody libraries are screened without blocking dominant epitopes, then the screening process is simpler and faster, but the epitopic coverage is biased towards dominant epitopes and fails to represent the full repertoire of epitopes
Solution Approach 1:
The method performs preliminary identification and characterization of dominant epitopes before the main screening process. This preliminary action allows the development of blocking reagents that can be applied in subsequent screening rounds, thereby preserving epitopic diversity information while maintaining screening efficiency through a structured multi-round approach.
Solution Approach 2:
The patent employs periodic action by conducting multiple sequential screening rounds with alternating objectives: some rounds focus on identifying dominant epitopes while others focus on discovering rare epitopes using blocking strategies. This periodic alternation ensures both screening efficiency and comprehensive epitopic coverage are achieved over the course of the complete screening process.
Data Source
AI summary
A method of broadening epitopic coverage of an antigen of interest, wherein a first sample of the antigen of interest is contacted with a first plurality of host cells collectively expressing a first library of antibodies. Host cells expressing antibodies that bind to the antigen are then collected from among the first plurality of host cells, and a composition is prepared comprising a polyclonal mixture of antibodies expressed by these host cells. A second sample of the antigen of interest is then contacted with an aliquot of the prepared composition and a second plurality of host cells collectively expressing a second library of antibodies. Host cells expressing antibodies that bind to the second sample of the antigen are then collected from among the second plurality of host cells.