Recombinant Monoclonal Antibody Libraries from Antigen-Binding B Cells
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for generating monoclonal antibodies are inefficient in surveying the entire antibody response induced by a particular immunogen, suffer from genetic instability, and have low production capacity, limiting the analysis of the vast immunoglobulin repertoire.
Innovation Solution
A method involving screening B cells to generate a library enriched in antigen-binding cells, amplifying cDNA to create immunoglobulin libraries, and expressing antibodies from VH and VL domains to identify recombinant antibodies capable of binding target antigens, utilizing high-throughput screening and evolution techniques to enhance diversity and affinity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If hybridoma methodology is used to generate monoclonal antibodies, then monoclonal antibodies can be produced, but the number of unique antibodies generated is limited to a few hundred per fusion
Solution Approach 1:
The patent segments the antibody generation process into separate B cell cloning and antibody production steps. Individual B cells are cloned and their antibody genes are sequenced, allowing for the identification and production of a much larger number of unique antibodies compared to traditional hybridoma fusion methods.
Solution Approach 2:
The patent uses molecular copying techniques to amplify and sequence the immunoglobulin genes from individual B cells. This allows for the efficient surveying and production of hundreds to thousands of unique antibody sequences from a single immunization event, far exceeding the limited output of hybridoma methods.
2Productivity
If hybridoma cultures are used to produce monoclonal antibodies, then antibody production is achieved, but genetic instability and low production capacity occur
Solution Approach 1:
The patent extracts the immunoglobulin genes from individual B cells and clones them into stable expression systems. This separates the antibody production function from the hybridoma cell line, allowing for genetic stability while maintaining high production capacity through controlled expression in suitable host cells.
Solution Approach 2:
Instead of maintaining long-term hybridoma cultures that suffer from genetic drift, the patent uses transient expression systems where antibodies are produced in stable, short-term cultures or in vitro translation systems, eliminating the genetic instability problem while maintaining production capacity.
3Quantity of substance
If the entire immunoglobulin repertoire is analyzed, then comprehensive antibody diversity can be surveyed, but the operation is expensive and time-consuming
Solution Approach 1:
The patent replaces manual, labor-intensive antibody analysis methods with automated high-throughput sequencing technology. This substitution allows for the rapid and cost-effective surveying of hundreds to thousands of unique antibody sequences from a single immunization, making comprehensive repertoire analysis practical and efficient.
Data Source
AI summary
The present invention relates to methods for efficiently generating recombinant monoclonal antibodies derived from B cells of a non-human host which has been immunochallenged with one or more target antigens. The methods comprise the steps of identifying and isolating B cell that bind to the antigen by FACS, and recombining and enriching for thousands of cells to create a B cell library. Related products and methods, such as methods of producing expression libraries, are also disclosed.