Simultaneous polyclonal antibody sequencing and epitope mapping by cryo electron microscopy and mass spectrometry.

ABSTRACT: Antibodies are a major component of adaptive immunity against invading pathogens. Here, we explore possibilities for an analytical approach to characterize the antigen-specific antibody repertoire directly from the secreted proteins in convalescent serum. This approach aims to perform simultaneous antibody sequencing and epitope mapping using a combination of single particle cryo-electron microscopy (cryoEM) and bottom-up proteomics techniques based on mass spectrometry (LC-MS/MS). We evaluate the performance of the deep-learning tool ModelAngelo in determining de novo antibody sequences directly from reconstructed 3D volumes of antibody-antigen complexes. We demonstrate that while map quality is a critical bottleneck, it is possible to sequence antibody variable domains from cryoEM recons

SUBMITTER: Schulte D 

PROVIDER: S-EPMC12017766 | biostudies-literature | 2025 Apr

REPOSITORIES: biostudies-literature