Mammalian cell display with automated oligo design and library assembly allows for rapid residue level conformational epitope mapping.

ABSTRACT: Precise epitope determination of therapeutic antibodies is of great value as it allows for further comprehension of mechanism of action, therapeutic responsiveness prediction, avoidance of unwanted cross reactivity, and vaccine design. The golden standard for discontinuous epitope determination is the laborious X-ray crystallography method. Here, we present a combinatorial method for rapid mapping of discontinuous epitopes by mammalian antigen display, eliminating the need for protein expression and purification. The method is facilitated by automated workflows and tailored software for antigen analysis and oligonucleotide design. These oligos are used in automated mutagenesis to generate an antigen receptor library displayed on mammalian cells for direct binding analysis by flow cytometry

SUBMITTER: Thalen NB 

PROVIDER: S-EPMC11222437 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature