Single B-Cell IgG Screening for Homogeneous Epitope Binding
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Solution Overview
Problem
Existing methods for producing antigen-binding proteins from a single B-lymphocyte often result in heterogeneous populations due to multiple VHDJH or VLJL recombination patterns, leading to proteins with undesired epitope specificity, avidity, and/or affinity.
Innovation Solution
A method involving cloning immunoglobulin heavy and light chains from a single B-cell, expressing them in host cells, screening for desired specificity and affinity, and optionally isolating the antigen-binding protein in eukaryotic cells to produce homogeneous antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If typical B-cell derived monoclonal antibody discovery techniques are used, then antigen-binding proteins can be produced from single B-cells, but heterogeneous antibody populations result due to multiple VHDJH or VLJL recombination patterns
Solution Approach 1:
The patent segments the antibody production process by separating the cloning of heavy chain and light chain genes from single B-cells into distinct steps, and further segments the screening process to evaluate each antibody variant individually for epitope specificity, allowing identification of homogeneous populations despite multiple recombination patterns
Solution Approach 2:
The patent changes the evaluation parameters by systematically assessing antigen-binding proteins not only for binding capability but specifically for epitope specificity, avidity, and affinity, allowing selection of homogeneous populations with desired characteristics from heterogeneous initial populations
2Adaptability or versatility
If multiple VHDJH or VLJL recombination patterns are present in single B-lymphocyte, then diverse antigen-binding proteins can be generated, but the antigen-binding region may not be specific to the epitope of interest
Solution Approach 1:
The patent implements feedback mechanisms through systematic screening and evaluation steps where antigen-binding proteins are tested for epitope specificity, and only those meeting specific criteria are selected for further production, creating a feedback loop that ensures precision despite diversity
Solution Approach 2:
The patent applies local quality by focusing evaluation on specific critical parameters (epitope specificity, avidity, affinity) rather than general binding capability, ensuring that the antigen-binding region has the desired local properties for the specific epitope of interest
3Productivity
If antigen-binding proteins are produced without systematic screening, then production speed is maintained, but accuracy of epitope binding properties decreases
Solution Approach 1:
The patent performs preliminary actions by establishing defined screening criteria and evaluation protocols before the actual production and screening process, allowing for efficient systematic evaluation that maintains productivity while ensuring precision through pre-planned assessment of epitope binding properties
Data Source
AI summary
The present disclosure provides a method of producing an antigen-binding protein (e.g., antigen-binding region, immunoglobulin, or antigen-binding fragment thereof), including cloning from a single B-cell at least one immunoglobulin heavy chain or an antigen-binding fragment thereof and at least one immunoglobulin light chain or an antigen-binding fragment thereof; producing at least one antigen-binding protein expressing host cell that expresses one or more of a single antigen-binding protein comprising one of the at least one immunoglobulin heavy chain or antigen-binding fragment thereof and one of the at least one immunoglobulin light chain or antigen-binding fragment thereof; screening/examining each of the at least one antigen-binding protein expressing host cell for the specificity, avidity, and/or affinity of the antigen-binding protein for a protein or peptide epitope of interest to find or select at least one antigen-binding protein; and expressing the at least one antigen-binding protein of interest in a eukaryotic cell.