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Precise epitope determination of therapeutic antibodies is of great value as it allows for further comprehension of the mechanism of action, therapeutic responsiveness prediction, avoidance of unwanted cross-reactivity, and vaccine design. Alanine scanning mutagenesis can provide an amino acid level resolution of a precise epitope for conformational epitopes. If the mutated antigen is presented on the surface of cells, high throughput screening is enabled without the need for protein expression and purification. Here we present a combinatorial method for rapid mapping of discontinuous epitopes by mammalian antigen display, facilitated by automated workflows for high throughput cloning and tailored software for antigen analysis and oligonucleotide design.

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Mammalian Cell Display with Automated Oligo Design and Library Assembly Allows for Rapid Residue Level Conformational Epitope Mapping

  • 实验方案

  • 发表日期：

    2025年

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  • Niklas Berndt Thalén,
  • Maximilian Karlander,
  • Magdalena Malm,
  • Johan Rockberg

摘要

Precise epitope determination of therapeutic antibodies is of great value as it allows for further comprehension of the mechanism of action, therapeutic responsiveness prediction, avoidance of unwanted cross-reactivity, and vaccine design. Alanine scanning mutagenesis can provide an amino acid level resolution of a precise epitope for conformational epitopes. If the mutated antigen is presented on the surface of cells, high throughput screening is enabled without the need for protein expression and purification. Here we present a combinatorial method for rapid mapping of discontinuous epitopes by mammalian antigen display, facilitated by automated workflows for high throughput cloning and tailored software for antigen analysis and oligonucleotide design.